Evaluation of kurtosis-corrected sound exposure level as a metric for predicting onset of hearing threshold shifts in harbor porpoises (Phocoena phocoena).

Application of a kurtosis correction to frequency-weighted sound exposure level (SEL) improved predictions of risk of hearing damage in humans and terrestrial mammals for sound exposures with different degrees of impulsiveness. To assess whether kurtosis corrections may lead to improved predictions for marine mammals, corrections were applied to temporary threshold shift (TTS) growth measurements for harbor porpoises (Phocoena phocoena) exposed to different sounds. Kurtosis-corrected frequency-weighted SEL predicted accurately the growth of low levels of TTS (TTS1-4 < 10 dB) for intermittent sounds with short (1-13 s) silence intervals but was not consistent with frequency-weighted SEL data for continuous sound exposures.

Noise of underwater explosions in the North Sea. A comparison of experimental data and model predictions.

An analysis is presented of sound measurements performed near two detonations of unexploded ordnance (UXO) in the North Sea, at distances ranging from 1.5 to 12 km. The charge masses of the detonations were 325 and 140 kg TNT equivalent. The objective of the measurements was to improve the accuracy of model predictions of the area where UXO detonations affect harbour porpoises in the North Sea. For the predictions, an explosion emission model is combined with a shallow-water propagation model. The prediction model was previously validated for distances up to 2 km. The measurements reported here allowed validation up to a distance of 12 km. The measured levels and spectra are well explained by the model calculations. The model results depend strongly on the sea sediment layering. The propagation of high-frequency components appears to be affected primarily by the silty top layer, while low-frequency components are affected also by deeper sandy layers. Measured and calculated noise levels are used to determine permanent-threshold-shift effect distances for harbour porpoises (Phocoena phocoena). Values ranging from 2 to 6 km are found for the two detonations.

Modeling potential masking of echolocating sperm whales exposed to continuous 1-2 kHz naval sonar.

Modern active sonar systems can (almost) continuously transmit and receive sound, which can lead to more masking of important sounds for marine mammals than conventional pulsed sonar systems transmitting at a much lower duty cycle. This study investigated the potential of 1-2 kHz active sonar to mask echolocation-based foraging of sperm whales by modeling their echolocation detection process. Continuous masking for an echolocating sperm whale facing a sonar was predicted for sonar sound pressure levels of 160 dB re 1 µPa2, with intermittent masking at levels of 120 dB re 1 µPa2, but model predictions strongly depended on the animal orientation, harmonic content of the sonar, click source level, and target strength of the prey. The masking model predicted lower masking potential of buzz clicks compared to regular clicks, even though the energy source level is much lower. For buzz clicks, the lower source level is compensated for by the reduced two-way propagation loss to nearby prey during buzzes. These results help to predict what types of behavioral changes could indicate masking in the wild. Several key knowledge gaps related to masking potential of sonar in echolocating odontocetes were identified that require further investigation to assess the significance of masking.

Predicting acoustic dose associated with marine mammal behavioural responses to sound as detected with fixed acoustic recorders and satellite tags.

To understand the consequences of underwater noise exposure for cetaceans, there is a need for assessments of behavioural responses over increased spatial and temporal scales. Bottom-moored acoustic recorders and satellite tags provide such long-term and large spatial coverage of behaviour compared to short-duration acoustic-recording tags. However, these tools result in a decreased resolution of data from which an animal response can be inferred, and no direct recording of the sound received at the animal. This study discusses the consequence of the decreased resolution of data from satellite tags and fixed acoustic recorders on the acoustic dose estimated by propagation modelling and presents a method for estimating the range of sound levels that animals observed with these methods have received. This problem is illustrated using experimental results obtained during controlled exposures of northern bottlenose whales (Hyperoodon ampullatus) exposed to naval sonar, carried out near Jan Mayen, Norway. It is shown that variability and uncertainties in the sound field, resulting from limited sampling of the acoustic environment, as well as decreased resolution in animal locations, can lead to quantifiable uncertainties in the estimated acoustic dose associated with the behavioural response (in this case avoidance and cessation of foraging).

Effect of towed array stability on instantaneous localization of marine mammals.

Reliable localization of marine mammals using towed arrays is often required for mitigation, population density estimates, and bioacoustics research. The accuracy of the range estimates using towed arrays is often not well quantified. Triangulation methods using multiple hydrophones allow for fast range estimates but are sensitive to the species type, location of the animal with respect to the array, sound propagation conditions, and array stability. A simple model is presented that is used to estimate the range accuracy of towed arrays for different vocalizations and is compared to measured range accuracies of sperm whale clicks recorded with a 15 m baseline towed array. The ranging performance is particularly sensitive to hydrophone position errors which are found to dominate. Hydrophone position errors could be estimated using heading sensors placed in the array and are taken into account in the model. A good agreement is found between the empirical range errors and theoretically predicted ones. Extrapolation of the model to other species suggests that species emitting high frequency clicks and calls can be localized from distances out to a few kilometers with a baseline of 15 m, but baleen whales transmitting low frequency calls require longer baselines to obtain range estimates.

History of circumcision, medical conditions, and sexual activity and risk of penile cancer.

BACKGROUND: Epidemiological evidence suggests lack of neonatal circumcision as the strongest risk factor for penile cancer, but the role of sexually transmitted diseases in the etiology of penile cancer has remained unclear. PURPOSE: To further clarify risk factors for penile cancer, we examined the role of circumcision, personal characteristics and habits (such as smoking), sexually transmitted diseases, past sexual activity, and medical conditions of the penis. METHODS: A population-based, case-control study was conducted in western Washington state and in the province of British Columbia. We interviewed 110 men with penile cancer diagnosed from January 1979 to July 1990 and 355 control subjects from the general population, frequency matched to case subjects on age and date of diagnosis. Tumor tissue from 67 case subjects was tested for human papillomavirus (HPV) DNA by polymerase chain reaction. Results of blood tests from 69 case subjects and 208 control subjects were available for study. STATISTICALLY SIGNIFICANT RESULTS: Relative to men circumcised at birth, the risk for penile cancer was 3.2 times greater among men who were never circumcised and 3.0 times greater among men who were circumcised after the neonatal period. For current smokers, the risk was 2.8 times that of men who never smoked. The risk among men reporting a history of genital warts was 5.9 times that of men reporting no such history. Of 67 tumors tested for HPV DNA, 49% were positive; the majority of these positive tumors (70%) were type 16, which has been associated with anogenital carcinoma. Relative risks (RRs) associated with a reported history of penile rash or penile tear were 9.4 and 3.9, respectively. Among men not circumcised at birth, RRs associated with presence of smegma and difficulty in retracting the foreskin were 2.1 and 3.5, respectively. Twenty-eight percent of case subjects, compared with only 10% of control subjects, reported 30 or more sexual partners, and men with HPV-positive tumors were more likely to report a greater number of sexual partners. CONCLUSIONS: These results suggest that the absence of neonatal circumcision and potential resulting complications are associated with penile cancer. Additionally, medical conditions of the penis, sexual activity, infection with HPV, and smoking may increase the risk for penile cancer. IMPLICATIONS: A larger study would allow examination of interrelationships of circumcision, infection with HPV, and smoking as risk factors.

Cofactors with human papillomavirus in a population-based study of vulvar cancer.

BACKGROUND: Human papillomavirus (HPV) has been previously associated with vulvar cancer. In a population-based study, we examined whether exposure to HPV, cigarette smoking, or herpes simplex virus 2 (HSV2) increases the risk of this cancer. METHODS: Incident cases of in situ (n = 400) and invasive (n = 110) squamous cell vulvar cancer diagnosed among women living in the Seattle area from 1980 through 1994 were identified. Serum samples were analyzed for antibodies against specific HPV types and HSV2. HPV DNA in tumor tissue was detected by means of the polymerase chain reaction. In most analyses, case subjects were compared with population-based control subjects (n = 1403). Relative risks of developing vulvar cancer were estimated by use of adjusted odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Increased risks of in situ or invasive vulvar cancer were associated with HPV16 seropositivity (ORs = 3.6 [95% CI = 2.6-4.8] and 2.8 [95% CI = 1.7-4.7], respectively), current cigarette smoking (ORs = 6.4 [95% CI = 4.4-9.3] and 3.0 [95% CI = 1.7-5.3], respectively), and HSV2 seropositivity (ORs = 1.9 [95% CI = 1.4-2.6] and 1.5 [95% CI = 0.9-2.6], respectively). When the analysis was restricted to HPV16 DNA-positive tumors (in situ or invasive), the OR associated with HPV16 seropositivity was 4.5 (95% CI = 3.0-6.8). The OR for vulvar cancer was 18.8 (95% CI = 11.9-29.8) among current smokers who were HPV16 seropositive in comparison with never smokers who were HPV16 seronegative. CONCLUSIONS: Current smoking, infection with HPV16, and infection with HSV2 are risk factors for vulvar cancer. Risk appears particularly strong among women who are both current smokers and HPV16 seropositive.

Oral cancer risk in relation to sexual history and evidence of human papillomavirus infection.

BACKGROUND: Experimental models and analyses of human tumors suggest that oncogenic, sexually transmittable human papillomaviruses (HPVs) are etiologic factors in the development of oral squamous cell carcinoma (SCC). We conducted a population-based, case-control study to determine whether the risk of this cancer is related to HPV infection and sexual history factors. METHODS: Case subjects (n = 284) were 18-65-year-old residents of three counties in western Washington State who were newly diagnosed with oral SCC from 1990 through 1995. Control subjects (n = 477) similar in age and sex were selected from the general population. Serum samples were tested for HPV type 16 capsid antibodies. Exfoliated oral tissue collected from case and control subjects and tumor tissue from case subjects were tested for HPV DNA. Odds ratios (ORs) were calculated after adjusting for age, sex, cigarette smoking, and alcohol consumption. RESULTS: Among males only, oral SCC risk increased with self-reported decreasing age at first intercourse, increasing number of sex partners, and a history of genital warts. Approximately 26% of the tumors in case subjects contained HPV DNA; 16.5% of the tumors contained HPV type 16 DNA. The prevalence of oncogenic HPV types in exfoliated oral tissue was similar in case and control subjects. The ORs for HPV type 16 capsid seropositivity were 2.3 (95% confidence interval [CI] = 1.6-3.3) for all oral SCCs and 6.8 (95% CI = 3.0-15.2) for oral SCCs containing HPV type 16 DNA. The joint association of cigarette smoking and HPV type 16 capsid seropositivity with oral SCC (OR = 8.5; 95% CI = 5.1-14.4) was stronger than predicted from the sum of individual associations with current smoking (OR = 3.2; 95% CI = 2.0-5.2) and seropositivity (OR = 1.7; 95% CI = 1.1-2.6). CONCLUSIONS: HPV type 16 infection may contribute to the development of a small proportion of oral SCCs in this population, most likely in combination with cigarette smoking.

First indications that northern bottlenose whales are sensitive to behavioural disturbance from anthropogenic noise.

Although northern bottlenose whales were the most heavily hunted beaked whale, we have little information about this species in its remote habitat of the North Atlantic Ocean. Underwater anthropogenic noise and disruption of their natural habitat may be major threats, given the sensitivity of other beaked whales to such noise disturbance. We attached dataloggers to 13 northern bottlenose whales and compared their natural sounds and movements to those of one individual exposed to escalating levels of 1-2 kHz upsweep naval sonar signals. At a received sound pressure level (SPL) of 98 dB re 1 µPa, the whale turned to approach the sound source, but at a received SPL of 107 dB re 1 µPa, the whale began moving in an unusually straight course and then made a near 180° turn away from the source, and performed the longest and deepest dive (94 min, 2339 m) recorded for this species. Animal movement parameters differed significantly from baseline for more than 7 h until the tag fell off 33-36 km away. No clicks were emitted during the response period, indicating cessation of normal echolocation-based foraging. A sharp decline in both acoustic and visual detections of conspecifics after exposure suggests other whales in the area responded similarly. Though more data are needed, our results indicate high sensitivity of this species to acoustic disturbance, with consequent risk from marine industrialization and naval activity.

HSV, CMV, and HPV in human neoplasia.

We are studying the role of sexually transmitted viruses in the development of human tumors. The persistence of herpes simplex virus, cytomegalovirus, and human papillomavirus nucleic acid sequences has been examined using cloned viral DNA sequences as probes. The relationship of the viruses to various stages in the progression of neoplasia is examined, with particular reference to the role of viral and/or cellular genes in the initiation, promotion, and maintenance of the neoplastic phenotype. The human tumors of major interest in this context are carcinomas of the cervix, vulva, and anus and Kaposi's sarcoma. The minimal fragment of HSV-2 DNA detected in cervical tumors is contained within a 656-bp sequence that can be used in transfection experiments to transform rodent cells in vitro to a malignant phenotype. However, neither this fragment nor any other is consistently retained in cervical tumors, suggesting that this viral DNA may initiate but not maintain the transformed phenotype.

Associations between human leukocyte antigen type and nasopharyngeal carcinoma in Caucasians in the United States.

A genetic component to nasopharyngeal carcinoma (NPC) has been suggested by associations of the malignancy with human leukocyte antigens (HLAs) in Southern Chinese populations, among which NPC is a major cancer. Data from other races are inconclusive. We have investigated associations between NPC and HLA antigens at the HLA-A, B, C, and DQ loci and alleles at the DRB1 locus in a population-based, multicenter investigation in the United States. Data from 82 cases and 140 controls are presented, making this the largest study population analyzing data from Caucasians to date. HLA frequencies from study cases were also compared with external control groups from the 11th International Histocompatibility Workshop and the National Marrow Donor Program. Logistic regression methods were used to investigate the effects of the joint occurrence of multiple HLA types and to assay for differences in HLA-associated risk in different age groups. A meta-analysis was undertaken to compare and summarize our results with previously published findings. The meta-analysis found a protective association with A2 antigen in non-Chinese [odds ratio (OR), 0.63; P < 0.001], a protective association with A11 across all races (OR, 0.54; P < 0.001), and an increased risk associated with B5 in Caucasians (OR, 2.81; P < 0.001). The present study also found independent associations, in a logistic regression model, between NPC and DRB1*1501 (OR, 0.33), DRB1*0405 (OR, 7.57), and Cw3 (OR, 0.42), although these data must be interpreted cautiously due to multiple-testing considerations. Associations were found to be more pronounced in younger patients for A2, A11, A28, B8, and B51.

The relationship of human papillomavirus-related cervical tumors to cigarette smoking, oral contraceptive use, and prior herpes simplex virus type 2 infection.

It has now been established that infection with human papillomavirus (HPV) is necessary for the development of most cervical cancers. HPV is not sufficient for the development of cancer. Other exposures or host factors are necessary for cancer to occur. As part of an ongoing, population-based case-control study of invasive cervical cancer, we investigated the role of cigarette smoking, oral contraceptive (OC) use, and herpes simplex virus type 2 (HSV-2) as potential cofactors with HPV in the development of cervical cancer. Residents of three counties in western Washington State who were diagnosed with invasive squamous cell cervical cancer (n = 314) from January 1986 through December 1992 were interviewed about their sexual, reproductive, contraceptive, and cigarette smoking histories. Similar information was obtained from control women identified through random digit dialing (n = 672). The sera from 206 cases and 522 controls were tested for both HPV 16 capsid antibodies and HSV-2 antibodies. PCR was used to test paraffin-embedded tumor tissues for the presence of HPV DNA types 6, 16, 18, 31, 33, 35, and 39. Women with cervical cancer were more likely to be current smokers at diagnosis than population controls [relative risk (RR), 2.5; 95% confidence interval (CI), 1.8-3.4]. The risk associated with smoking was present to a similar extent among women positive and negative for HPV as measured by HPV 16 capsid antibodies and HPV DNA in the tumor tissue (cases). OC use was only important if first use was at an early age, particularly ages < or = 17 years (RR, 2.3; 95% CI, 1.4-3.8). There was only a slight risk for cervical cancer associated with antibodies to HSV-2 (RR, 1.2; 95% CI, 0.9-1.7). However, when we stratified by markers of HPV exposure, we found a significant increase in risk associated with HSV-2 among women negative for HPV 16 antibodies (RR, 2.0; 95% CI, 1.3-3.0), which was strengthened when we confined our analysis to cases whose tumors were HPV DNA negative (RR, 3.6; 95% CI, 1.6-8.0). There was no indication that cigarette smoking, OC use, or HSV-2 infection influence the ability of HPV infection to cause invasive cervical cancer. OC use may only be important in the etiology of invasive squamous cell cervical tumors if the use occurs at a critical time in the development of a woman's reproductive tract, at ages < or = 17 years. The majority of risk associated with HSV-2 was confined to HPV-negative tumors, indicating a possible separate pathway to disease that may account for 5-10% of invasive cervical cancers.

Immediate early gene expression during morphine withdrawal.

The expression of immediate early genes (IEG)s c-fos, c-jun and zif/268 was studied during naloxone-precipitated morphine withdrawal in various organs of the rat. Dependence was induced over a period of 6 days by a graded regimen of 6-hourly injections. Northern analysis revealed peak expression of all IEGs occurred in the forebrain plus cerebellum at 20 min and at 60 min in the brain stem following morphine withdrawal. Increased levels of c-fos and c-jun mRNA were observed in the spinal cord at 40 min of morphine withdrawal. An increase in c-fos and c-jun but not zif/268 mRNAs was seen in the jejunum between 20 and 60 min. Elevated levels of the IEG protein products in the cerebral cortex, hippocampus, thalamus, cerebellum, brain stem and spinal cord were observed at 60 min following morphine withdrawal. These data emphasize the temporal and spatial variation in IEG expression in different tissues during opiate withdrawal.

Detection of viral DNA and RNA by in situ hybridization.

Using cloned restriction endonuclease fragments of Herpes simplex virus (HSV), human papillomavirus (HPV), and cytomegalovirus (CMV) DNA as probes, viral DNA and RNA sequences have been detected in human tissues. The probes were labeled either with a radioactive isotope, for subsequent detection by autoradiography, or with biotin. This latter technique has been successfully used to visualize HPV DNA in tissues that have been fixed in formalin and embedded in paraffin, and is therefore of value in retrospective studies of histological specimens. HPV DNA was detected under non-stringent conditions (Tm = -42 degrees C) with heterologous probes in plantar and common warts, laryngeal papillomas, and anogenital condylomas. The specific type of HPV was established using stringent hybridization conditions (Tm = - 17 degrees C). Results from these and from malignant tissues show the distribution and localization of HSV and HPV RNA and DNA sequences in malignancies of squamous cell origin in the anogenital region. Both HSV and HPV DNA sequences have occasionally been detected in the same tumor, providing a further impetus to test the hypothesis that an initiator-promoter relationship might involve these common human viruses in the development of some tumors.

Sequential increase in Egr-1 and AP-1 DNA binding activity in the dentate gyrus following the induction of long-term potentiation.

Establishment of long-term potentiation (LTP) at perforant path synapses is highly correlated with increased expression of Egr and AP-1 transcription factors in rat dentate gyrus granule cells. We have investigated whether increased transcription factor levels are reflected in increased transcription factor activity by assessing Egr and AP-1 DNA binding activity using gel shift assays. LTP produced an increase in binding to the Egr element, which was NMDA receptor-dependent and correlated closely with our previously reported increase in Egr-1 (zif/268) protein levels. Supershift analysis confirmed involvement of Egr-1, but not Egr-2 in the DNA binding activity. AP-1 DNA binding was also rapidly elevated in parallel with protein levels, however, the peak increase in activity was delayed until 4 h, a time point when we have previously shown that only jun-D protein was elevated. These data indicate that binding of Egr-1 and AP-1 to their response elements is increased in two phases. This may result in activation of distinct banks of target genes which contribute to the establishment of persistent LTP.

Egr transcription factors in the nervous system.

The Egr proteins, Egr-1, Egr-2, Egr-3 and Egr-4, are closely related members of a subclass of immediate early gene-encoded, inducible transcription factors. They share a highly homologous DNA-binding domain which recognises an identical DNA response element. In addition, they have several less-well conserved structural features in common. As immediate early proteins, the Egr transcription factors are rapidly induced by diverse extracellular stimuli within the nervous system in a discretely controlled manner. The basal expression of the Egr proteins in the developing and adult rat brain and the induction of Egr proteins by neurotransmitter analogue stimulation, physiological mimetic and brain injury paradigms is reviewed. We review evidence indicating that Egr proteins are subject to tight differential control through diverse mechanisms at several levels of regulation. These include transcriptional, translational and post-translational (including glycosylation, phosphorylation and redox) mechanisms and protein-protein interaction. Ultimately the differentially co-ordinated Egr response may lead to discrete effects on target gene expression. Some of the known target genes of Egr proteins and functions of the Egr proteins in different cell types are also highlighted. Future directions for research into the control and function of the different Egr proteins are also explored.

Glutamate-mediated transmission, alcohol, and alcoholism.

Glutamate-mediated neurotransmission may be involved in the range of adaptive changes in brain which occur after ethanol administration in laboratory animals, and in chronic alcoholism in human cases. Excitatory amino acid transmission is modulated by a complex system of receptors and other effectors, the efficacy of which can be profoundly affected by altered gene or protein expression. Local variations in receptor composition may underlie intrinsic regional variations in susceptibility to pathological change. Equally, ethanol use and abuse may bring about alterations in receptor subunit expression as the essence of the adaptive response. Such considerations may underlie the regional localization characteristic of the pathogenesis of alcoholic brain damage, or they may form part of the homeostatic change that constitutes the neural substrate for alcohol dependence.

Propagation and primary isolation of JCV and BKV in urinary epithelial cell cultures.

Human papovavirus JC, previously passaged in amnion cells, produced a cytopathic effect in urine-derived epithelial cells, and virus-specific antigens were demonstrable by indirect immunofluorescence. The hemagglutinating titer of JCV purified from infected cell cultures was generally 100- to 1000-fold higher than the amount of viral hemagglutinin used to initiate infection. Amnion-passaged JCV was readily adapted to growth in urine-derived epithelial cells. The prototype strain of human papovavirus BK, adapted to growth in human embryonic lung cells, also productively infected urine-derived epithelial cells. Primary human fetal glial cells and urine-derived cells were used in parallel experiments for primary isolation of JCV from diseased brain tissue. For this purpose, primary human fetal glial cells were more sensitive than urine-derived epithelial cells. Primary isolations of JCV and BKV from urine sediments of transplant patients and a normal male were made in urine-derived cells. Two renal transplant patients were identified as simultaneously excreting JCV and BKV. Both JCV and BKV genomes were molecularly cloned from one urine specimen of a double excretor. Although direct comparisons between primary human fetal glial and urine-derived epithelial cells were not made, it appears that the latter may be more suitable for primary isolation of JCV from urine.

Methods for diagnosing papillomavirus infection.

The morphology of the lesion and the site in which the lesion is found are the initial clues in classifying papillomavirus-induced neoplasia. Human papillomavirus (HPV) types have limited site-specificity and differ in their association with benign or malignant neoplastic development. Cytopathology, electron microscopy, antigen detection and molecular hybridization all play a role in the armamentarium of diagnostic methods. Although nitrocellulose blotting procedures provide the most accurate and sensitive method for detecting and characterizing viral nucleic acid sequences, recent improvements in cytological hybridization methods allow for rapid detection of virus and analysis of HPV type directly in biopsied tissue and in cervical smears. In particular, these in situ hybridization procedures facilitate retrospective studies of stored specimens.