Detection of high-risk HPV DNA in semen and its association with the quality of semen.

The effects of seminal high-risk human papillomavirus (HPV) DNA were assessed on the quality of semen. Semen samples of 65 men participating in the ongoing Finnish HPV Family Study were collected. Semen analyses were done by the guidelines of the Nordic Association for Andrology. HPV DNA was detected by nested polymerase chain reaction and confirmed by Southern blot hybridization for high-risk types. Altogether, 10/65 men (15.4%) had high-risk HPV DNA positive semen sample. Seminal high-risk HPV DNA did not affect semen volume, sperm concentration, motility and vitality of spermatozoa. However, semen pH was borderline lower in HPV DNA positive than negative samples (7.4 vs 7.5). Neither oligo- nor asthenozoospermia was associated with seminal HPV DNA. In conclusion, seminal high-risk HPV DNA was detected in 15% of men. It did not affect the semen analysis, except semen pH by borderline significance. Sperm donors have not been tested for HPV infections, sperm washing does not seem to eliminate the risk of HPV transmission and the consequences of HPV in the semen are at present unknown.

Association of syndecan-1 with tumor grade and histology in primary invasive cervical carcinoma.

OBJECTIVES: The expression of syndecan-1, a cell surface heparan sulfate proteoglycan, is reduced during malignant transformation of squamous cells. Studies on squamous cell carcinoma of the head and neck have shown that syndecan-1-positive tumors are associated with longer overall and recurrence-free survival. The purpose of this study was to analyze syndecan-1 expression in invasive cervical carcinoma and to examine the association of syndecan-1 expression with prognostic factors and overall survival. METHODS: The study population consisted of 124 patients treated for primary invasive carcinoma of the uterine cervix at the Turku University Central Hospital during the years 1970-1988. The material consisted of 102 (82.3%) squamous cell carcinomas, 16 (12.9%) adenocarcinomas and 1 (0.8%) adenosquamous carcinoma, 1 (0.8%) small cell carcinoma, 1 (0. 8%) adenoid basal carcinoma, 1 (0.8%) carcinosarcoma, and 2 (1.6%) unclassified cervical carcinomas. Syndecan-1 expression was determined on paraffin-embedded tissue blocks using a human syndecan-1-specific monoclonal antibody B-B4 and immunohistochemistry. The expression of syndecan-1 was classified according to staining intensity as well as the percentage of positively stained tumor cells. RESULTS: Staining intensity was strong in 44 (36%) samples, while 24 (19%) specimens remained syndecan-1-negative. In 49 (40%) samples, the percentage of syndecan-1-positive cells was >/=90%. Syndecan-1 expression, as determined by >/=50% positively stained tumor cells, was associated with the grade of differentiation (P = 0.03) and squamous histology (P < 0.001), but was not associated with clinical stage (P = 0.16) or disease-free survival (P = 0.86). Age (P = 0.003) and clinical stage (P < 0.001) were significant prognostic factors, but syndecan-1 expression determined neither by percentage of positively stained tumor cells nor by staining intensity was associated with the outcome. CONCLUSIONS: In cervical carcinoma syndecan-1 is associated with histological differentiation grade and squamous histology, but does not predict clinical outcome.

Tissue-separating capacity of growth cartilages.

The tissue-separating capacity of chondral structures has been debated for more than 30 years, and one aspect that has particularly been questioned is whether the secondary cartilage of the mandibular condyle is comparable to primary growth cartilage, e.g. the epiphyseal growth plate. The present report summarizes information gained by using a specific interosseal transplantation method. These findings lead to the conclusion that all the structures examined, i.e. the proximal epiphyseal cartilage of the tibia, the cartilage of the costochondral junction of the ribs, the basicranial synchondroses, the medial cartilage of the clavicle and the mandibular condyle, have the capacity to separate adjoining skeletal structures. The changes induced by the transplanted structures in the recipient area vary, however, suggesting a hierarchial arrangement of cartilages with regard to their tissue-separating capacity. It is suggested that the tissue-separating capacity is a basic phenomenon in the function of growth, not only of primary growth cartilages, but of secondary cartilages as well.

Abnormal craniofacial growth and early mandibular osteoarthritis in mice harbouring a mutant type II collagen transgene.

Skull morphology and histology in the heterozygous offspring of a transgenic founder mouse Del1, harbouring 6 copies of deletion mutation in Col2a1 gene, were compared with those in normal siblings. On visual observation and roentgenocephalometric examination the heads of heterozygous Del1 mice were smaller than normal. Histologically the sizes of cartilaginous structures of the cranial base were reduced. Severe defects were seen in the temporomandibular joint as progressive osteoarthritic lesions. These observations elucidate the relationship between the genotype and phenotype and demonstrate that heterozygous Del1 mice are a useful model for studies on a genetic disturbance where 'clinical' manifestations are not evident until adult age.

PAP smear after radiation therapy for cervical carcinoma.

Vaginal PAP smear is frequently used for the follow-up of cervical carcinoma after primary therapy. Irradiation induced atypia can interfere with cytological analysis and thus detection of a local recurrence, or simulate malignant atypia and cause unnecessary suspicion of recurrence. In this retrospective study we evaluated the reliability of cytological analysis and the reported frequency of irradiation induced atypia after radiotherapy. Eighty-nine patients treated for cervical carcinoma at Turku University Central Hospital during the years 1970-88 were included in the study. During the median follow-up of 34 months a total of 697 PAP smears were taken with a median of 7.8 samples per patient. During the follow-up 44 (50%) patients had a recurrent disease, which was local in 17 (39%) cases. Nine out of 12 PAP smears taken 0-60 days before detection of a local recurrence showed class III-V cellular atypia. However, three PAP smears showed class I-II, and were therefore false negative. The rate of false positive samples was only 3%. In 567 PAP smears irradiation induced atypia was indicated as present/not present (+/-) and it was positive in 89 (16%) samples. The detection rate was considerably higher (75%) in class II samples than in rest of the material. Irradiation induced atypia was detected in 28% of the PAP smears taken during the first four months after radiotherapy and the rate decreased thereafter. Cytological analysis of vaginal PAP smear was a reliable indicator of recurrence in most cases and is a valuable tool for the detection of local recurrence of cervical carcinoma after primary radiotherapy.

Ocular abnormalities in transgenic mice harboring mutations in the type II collagen gene.

PURPOSE: To characterize the morphological changes in the eyes of transgenic mice harboring different mutations in type II collagen gene to elucidate the function of this collagen in the eye, and to find out whether these animals could function as models for the human arthro-ophthalmopathies of the Kniest, Stickler and Wagner types. METHODS: Three genetically engineered mouse lines representing two types of mutations in the triple-helical domain of type II collagen and their nontransgenic littermates used as controls were analyzed on day 18.5 embryonic development. After genotyping by polymerase chain reaction (PCR) and Southern hybridization the embryos were prepared for routine histology. Polarization microscopy was done on hyaluronidase-treated sections. RESULTS: Histological analysis revealed several genotype-dependent abnormalities in the eyes of the transgenic mice. Most striking changes were observed in the vitreous architecture; in one line of mice the vitreous was tightly packed in the posterior region of the vitreous space with thick fibrils, empty cavities and dense membrane-like material. The other mutation resulted in reduced filament density of the vitreous. In the most severely affected phenotype the internal limiting membrane was detached from the retinal layers and was markedly thickened, and the posterior lens capsule was thickened. The anterior chamber was shallow or absent in all transgenic lines but was well formed in the normal animals. Changes were also observed in the lens, corneal and scleral structures. CONCLUSIONS: The ocular changes observed in transgenic mice harboring mutations in type II collagen gene show similarities to the human ocular findings in Kniest dysplasia, and in Stickler and Wagner syndromes. We therefore propose that these animals could serve as models for systematic analysis of vitreoretinal degeneration and other abnormalities, as seen in these syndromes.

Abnormalities in secondary cartilages in four lines of transgenic mice harboring two different types of mutations in the cartilage-specific type II collagen gene.

A histological analysis was performed on two secondary cartilages, the mandibular condyle and the medial end of clavicle, in transgenic mice harboring two different types of mutations in the cartilage-specific type II collagen gene. Considerable differences were observed in the maturation zone of the chondrocytes and the hypertrophic cells of the growth regions of the two secondary cartilages examined between the transgenic mice and their transgene-negative littermates, which served as controls. Looseness of the perichondrium/periosteum was a distinct feature seen in both mutations. However, phenotypic consequences of the mutations in secondary cartilages were less severe than those in primary cartilages. We propose that the differences between primary and secondary cartilages are due to differences in their origin, mode of growth, architecture and behavior under extrinsic factors.

Abnormal craniofacial morphology and cartilage structure in transgenic mice harboring a Gly --> Cys mutation in the cartilage-specific type II collagen gene.

Gross morphology and histology of the craniofacial complex was studied in the offspring of two transgenic founder mice, Gly85-1 and Gly85-3, carrying several copies of a mouse type II collagen transgene that causes a single amino acid substitution (Gly-->Cys) at position 85 of the triple helix. The newborn transgenic mice had a short snout and mandible, a protruding tongue, a cleft of the secondary palate with the tongue situated between the shelves, and a doming cranial vault. Radiological examination revealed that the cranial base of the transgenic mice was shorter and its posterior part downward bent; in addition both the palate and the cribriform plate were less extended in relation to the cranial base than in the controls. Histologically the midline cartilaginous structures were composed of densely packed enlarged chondrocytes in a reduced extracellular matrix containing abnormally thick collagen fibrils. With the exception of the zone of hypertrophic chondrocytes the matrix also showed a loss of glycosaminoglycans. The cellular architecture of the basicranial synchondroses was disorganized, and the nasocerebrally oriented collagen fibrils formed unevenly distributed aggregates. The craniofacial morphology described here for the Gly85 mice shares features typical for other mouse mutations causing short limbed dwarfism. The observations indicate that defective cartilage production causes disproportionate craniofacial growth. Transgenic mice with specific mutations in cartilage-specific genes should therefore be useful for elucidating the complex mechanisms involved in determining the craniofacial growth.

Chondrodysplasia in transgenic mice harboring a 15-amino acid deletion in the triple helical domain of pro alpha 1(II) collagen chain.

We have generated transgenic mice by microinjection of a 39-kb mouse pro alpha 1(II) collagen gene construct containing a deletion of exon 7 and intron 7. This mutation was expected to disturb the assembly and processing of the homotrimeric type II collagen molecule in cartilage. Expression of transgene mRNA at levels equivalent or higher than the endogenous mRNA in the offspring of two founder animals resulted in a severe chondrodysplastic phenotype with short limbs, hypoplastic thorax, abnormal craniofacial development, and other skeletal deformities. The affected pups died at birth due to respiratory distress. Light microscopy of epiphyseal growth plates of transgenic pups demonstrated a marked reduction in cartilaginous extracellular matrix and disruption of the normal organization of the growth plate. The zone of proliferating chondrocytes was greatly reduced whereas the zone of hypertrophic chondrocytes was markedly increased extending deep into the diaphysis suggestive of a defect in endochondral ossification. Electron microscopic examination revealed chondrocytes with extended RER, a very severe reduction in the amount of cartilage collagen fibrils, and abnormalities in their structure. We postulate that the deletion in the alpha 1(II) collagen acts as a dominant negative mutation disrupting the assembly and secretion of type II collagen molecules. The consequences of the mutation include interference with normal endochondral ossification. These mice constitute a valuable model to study the mechanisms underlying human chondrodysplasias and normal bone formation.

Growth potential of the rat clavicle.

An experiment was performed to determine whether the clavicle, which is capped with secondary cartilage at both ends, is endowed with a tissue-separating quality or is only adaptive to forces exerted by the surrounding structures. The medial end of the clavicle from 10-day-old male rats was transplanted across the interparietal suture into an opening in the adjoining parietal bones corresponding to the outline of the transplant. The transverse dimensions of the neurocranium when measured at 25 and 35 days of age were greater in the animals with transplants than in the untreated controls. Histologic examination showed that although the cartilaginous component of the transplants was reduced, the zonal arrangements was comparable to that in situ. It was concluded that the medial end of the clavicle has a potential for separating skeletal components and that preservation of the basic structure is dependent on mechanical resistance.