Short-time repeat high-risk HPV testing by self-sampling for screening of cervical cancer.

BACKGROUND: Testing for high-risk human papillomavirus (HPV) in primary screening for cervical cancer is considered more sensitive, but less specific, in comparison with Pap-smear cytology. Women with persistent HPV infections have a higher risk of developing cervical intraepithelial neoplasia 2+ (CIN2+) lesions. This study was performed to evaluate the gain in specificity for detection of histologically confirmed CIN2+ lesions achieved by short-time repeat testing for high-risk HPV in women aged 30-65 years, with the primary sample for HPV analysis taken by self-sampling. METHODS: A total of 8000 women in Uppsala County, aged 30-65 years, who had not attended organised screening for 6 years or longer, were offered self-sampling of vaginal fluid at home and the samples sent for HPV typing. Of these, 8% (669) were not possible to contact or had performed hysterectomy. Women positive for high-risk HPV in the self-sampling test were invited for a follow-up HPV test and a cervical biopsy on average 3 months after the initial HPV test. RESULTS: In all, 39% (2850/7331) of invited women chose to perform self-sampling of vaginal fluid at home. High-risk HPV infection was found in 6.6% (188) of the women. In all, 89% of the women testing HPV positive performed a follow-up examination, on average 2.7 months, after the first test and 59% of these women were HPV positive in the follow-up test. The prevalence of CIN2+ lesions in women with an initial HPV-positive test was 23% (95% CI 18-30%) and in women with two consecutive HPV-positive tests was 41% (95% CI 31-51%). In women with two positive HPV tests, the prevalence of CIN2+ lesions varied from 49% in women at age 30-39 years to 24% in women at age 50-65 years. Short-time repeat HPV testing increased the specificity for detection of CIN2+ lesions from about 94.2% to 97.8%. The most prevalent HPV types were HPV16 (32%), followed by HPV18/45 (19%) and HPV 33/52/58 (19%). CONCLUSION: The short-time persistence of high-risk HPV infection in this age group was about 60%. Repeat testing for high-risk HPV using self-sampling of vaginal fluid can be used to increase the specificity in the screening for cervical cancer in women aged 30-65 years.

A primary linkage map of the porcine genome reveals a low rate of genetic recombination.

A comprehensive genetic linkage map of the porcine genome has been developed by typing 128 genetic markers in a cross between the European Wild Boar and a domestic breed (Large White). The marker set includes 68 polymerase chain reaction-formatted microsatellites, 60 anchored reference markers informative for comparative mapping and 47 markers which have been physically assigned by in situ hybridization. Novel multipoint assignments are provided for 54 of the markers. The map covers about 1800 cM, and the average spacing between markers is 11 cM. We used the map data to estimate the genome size in pigs, thereby addressing the total recombination distance in a third mammalian species. A sex-average genome length of 1873 +/- 139 cM was obtained by comparing the recombinational and physical distances in defined regions of the genome. This is strikingly different from the length of the human genome (3800-4000 cM) and is more similar to the mouse estimate (1600 cM). The recombination rate in females was significantly higher than in males.

Sex steroid receptors in human myometrium and fibroids: changes during the menstrual cycle and gonadotropin-releasing hormone treatment.

The content of estrogen and progesterone receptors (ER, PR) is higher in fibroid tissue than in homologous myometrium, and both receptors seem to be regulated by the levels of circulating sex steroids. Myometrial and fibroid tissues were recovered from women undergoing gynecological operations during different phases of the menstrual cycle and during treatment with an analogue of GnRH (GnRHa). Contents of ER and PR in the tissue cytosol were determined by enzyme immunoassay. The ER levels were significantly higher in fibroid tissue than in homologous myometrium in all the endocrine conditions. During the secretory phase, when luteal progesterone production is prominent, the ER levels in the myometrium and fibroids were lower than during the proliferative phase. During GnRHa treatment, the ER levels in both tissues were similar to those in the proliferative phase but significantly higher than in the secretory phase. The PR levels were also significantly higher in fibroids than in myometrium in all the different endocrine conditions. In both tissues, the PR levels were lower in the secretory phase and during GnRHa treatment, compared with the proliferative phase. Our data suggest that, in these categories of women, both ER and PR are overexpressed in fibroid tissue. Apparently, high progesterone levels down-regulate the ER in both fibroids and myometrium, whereas estrogen mediates the up-regulation of the PR during the proliferative phase. Increased knowledge about the mechanisms by which sex steroids regulate their own receptors in uterine tissues might provide a basis for development of new treatment strategies for women with fibroid disease.

Benign ovarian teratomas. An analysis of their cellular origin.

To determine the cellular origin of benign ovarian teratomas with a 46,XX chromosome constitution, DNA markers recognizing restriction fragment length polymorphisms (RFLPs) were hybridized to DNA from six teratomas and their hosts. DNA markers heterozygous in the host were completely heterozygous in two of the teratomas. The remaining four showed a mixture of homozygosity and heterozygosity. These results suggests that most of the analyzed benign ovarian teratomas arose from germ cells after the first meiotic division by failure of meiosis II. Teratomas heterozygous for all tested markers may arise from failure of meiosis I. In addition, 21 cases were karyotyped and analyzed for centromeric chromosome markers to study the mechanism by which they were generated. Three of these tumors were homozygous when the host was heterozygous and therefore resulted from a failure of meiosis II or duplication of a mature ovum. Three cases were heterozygous for the centromeric chromosomal marker like the host and therefore probably originate from a premeiotic cell or a cell in which meiosis I has failed. One ovarian teratoma had an aberrant karyotype 47,XX,+8.

Synaptonemal Complex Analysis of Spermatocytes in Hybrids of Silver Fox and Blue Fox.

Investigations of male meiosis in silver fox x blue fox hybrids have revealed meiotic arrest at the first prophase stage. Synaptonemal complex analysis using light and electron microscopy demonstrated the occurrence of multivalents, bivalentlike structures, and unpaired axes. We conclude that the sterility of male hybrid foxes probably is due to pairing problems of chromosomes caused by extensive karyotypical differences of the two species, resulting in unpaired chromosomes, chromosome segments, and broken chromosomes.

Simultaneous detection of viability and sex of bovine spermatozoa.

The viability and sex of bovine spermatozoa were simultaneously evaluated. After viability and acrosome staining with trypan blue/Giemsa, only live spermatozoa became decondensed by a modified papain-dithiothreitol method. Owing to this specific effect, live sperm heads were easily distinguished by their enlarged size and dark violet colour from small, light blue dead sperm heads. In the same sperm sample, X- and Y-chromosome-bearing sperm were distinguished by their fluorescent signal, using fluorescence in situ hybridization (FISH) with an XY paint set and 4,6-diamino-2-phenylindole counterstaining. The combined staining provides a method for morphological and viability evaluation before FISH and permits identification of the proportions of X- and Y-chromosome-containing live spermatozoa in a semen sample. However, only 25% of the undecondensed dead sperm express signals allowing detection of the sex of the chromosome. The method may be an effective tool in evaluating sex-oriented semen samples.

Head area measurements of dead, live, X- and Y-bearing bovine spermatozoa.

The head area of bull spermatozoa was measured after viability and acrosome staining using trypan blue and Giemsa stains, followed by X- and Y-chromosome-specific fluorescence in situ hybridisation (FISH). The former staining made possible the categorisation of cells according to morphology and membrane integrity, whereas the latter allowed distinction of spermatozoa bearing X- and Y-chromosomes. Individual spermatozoa could be followed during the consecutive steps of staining, measurement and FISH. Using a high-resolution digital imaging system and measurement software, the head area of more than 3000 cells of five bulls was determined precisely. In all bulls, morphologically normal, viable cells with intact acrosomes were significantly smaller than dead cells with damaged acrosomes. No significant difference in the head area between X- and Y-chromosome-bearing viable, acrosome-intact spermatozoa was found in individual bulls. However, significant between-bull differences were detected in all cell categories.

A pyrolytic carbon film electrode for voltammetry-III Application to anodic-stripping voltammetry.

The potential applicability of a pyrolytic carbon film electrode in the differential-pulse anodic-stripping voltammetric determination of cadmium and lead in sea-water is demonstrated. The performance at the 10(-10)M level is compared with that of a satisfactory glassy-carbon electrode. The two types of electrode display comparable behaviour in anodic-stripping voltammetry, but the pyrolytic carbon film electrode needs less pretreatment.

Determination of mercury in freeze-dried muscle samples of pike, cod and perch using an ASV-technique.

Determination of mercury in fish muscle samples can be made by an anodic stripping voltammetric technique (ASV) using a gold disc as the working electrode. Prior to analysis deep-frozen fish were thawed and then dissected using quartz scalpels. The freeze-dried samples were homogenised by the brittle fracture technique. Two wet digestion procedures were investigated, namely the HNO3/HCIO4 and the HNO3/H2SO4 system, and both were found to be useful. In order to complete the oxidation, the sample had to be UV-irradiated. The stripping procedure can be done in either a new medium - 0.1 M HCIO4 and 2.5 mM HCl - or directly in the sample solution. Using HNO3 and H2SO4 as oxidising agents and subsequently stripping the mercury directly in the sample solution is recommended, as the analytical procedure will be simpler and more time-saving. The concentrations obtained for pike, cod and perch, caught at a near-shore Baltic Sea station off the Forsmark nuclear power plant in Sweden, were 19.6, 0.84 and 5.1 (micrograms/g dry weight), respectively. On comparison with results obtained from neutron activation analysis, good agreement was found.

60,XY/60,XX chimerism in the germ cell line of mature bulls born in heterosexual twinning.

According to present knowledge there is a germ cell chimerism (XY/XX) in young bulls born in heterosexual twinning due to exchange of primordial germ cells in embryonic life. These germ cells were believed to have been eliminated in the young bull. Two-color fluorescence in situ hybridization (FISH) identification of the sex chromosomes by biotinylated and digoxygenin labeled probes have been used. The material consisted of three bulls born in heterosexual twinning. The results obtained indicated that even mature bulls (more than two years old) demonstrate spermatogonial chimerism. Several authors state that the bulls with blood cell chimerism, originating from dizygous twinning, are characterized by decreased fertility. Changes of the sex ratio of offspring due to proliferation of the female cells have also been proposed. The present observations should give a renewed interest in checking the possibility of survival and differentiation of germ cells from the female partner in the germ cell lines.

Embryo quality and andrological study of two subfertile bulls versus five control bulls with normal fertility.

Andrological studies and embryo morphology evaluation of superovulated cows were performed on 2 randomly selected subfertile dairy bulls whose semen was used for artificial insemination and on 5 control bulls with normal fertility. Neither sperm motility studies, nor sperm morphology or testicular measurements differed between the subfertile and the control bulls. Altogether 315 ova were recovered from 41 superovulated cows inseminated with semen collected from either the subfertile or the normal control bulls. The spermatozoa of one of the 2 subfertile bulls was shown to have a decreased ability to fertilize superovulated ova, while the other subfertile animal, the bull with the lowest no-return rate, was found by chromosome analysis to have a reciprocal translocation (60, XY, rcp 20:24), causing embryonic death. We suggest that subfertile bulls should not be used in commercial embryo transfer programs nor in artificial insemination and that andrological studies on subfertile bulls with good sperm motility should include evaluation of 6- to 7-day-old ova from superovulated cows to determine if the fertilization rate is normal or impaired. A chromosome analysis should also be performed when a subjertile bull has a normal fertilization rate of ova.

Reproductive disorders in 10 domestic male cats.

This study describes 10 tomcats with different reproductive disorders. Two of the cats had abnormal sex chromosomes; one was a tortoiseshell and white Cornish rex, while the other was a brown Burmese. The other eight cats were diagnosed as having testicular hypoplasia, diphallos in combination with unilateral cryptorchidism, a persistent penile frenulum, retrograde ejaculation, temporary oligozoospermia, teratozoospermia, azoospermia and congenital poor libido. For the cat with a persistent penile frenulum, and the cat with a temporary oligozoospermia, the prognosis for successful reproduction was considered favourable. By contrast it was considered unlikely that the cats with chromosomal abnormalities, testicular hypoplasia, diphallos, retrograde ejaculation, teratozoospermia and azoospermia would be able to produce offspring.

Temporal trends of HLA genotype frequencies of type 1 diabetes patients in Sweden from 1986 to 2005 suggest altered risk.

The aim of this study was to compare the frequency of human leukocyte antigen (HLA) genotypes in 1-18-year-old patients with type 1 diabetes newly diagnosed in 1986-1987 (n = 430), 1996-2000 (n = 342) and in 2003-2005 (n = 171). We tested the hypothesis that the HLA DQ genotype distribution changes over time. Swedish type 1 diabetes patients and controls were typed for HLA using polymerase chain reaction amplification and allele specific probes for DQ A1* and B1* alleles. The most common type 1 diabetes HLA DQA1*-B1*genotype 0501-0201/0301-0302 was 36% (153/430) in 1986-1987 and 37% (127/342) in 1996-2000, but decreased to 19% (33/171) in 2003-2005 (P \ 0.0001). The 0501-0201/0501-0201 genotype increased from 1% in 1986-1987 to 7% in 1996-2000 (P = 0.0047) and to 5% in 2003-2005 (P > 0.05). This study in 1-18-year-old Swedish type 1 diabetes patients supports the notion that there is a temporal change in HLA risk.