SOUTHERN BLOT ANALYSIS OF SKIN BIOPSIES FOR HUMAN PAPILLOMAVIRUS DNA: RENAL ALLOGRAFT RECIPIENTS IN SOUTH-EASTERN QUEENSLAND

The 104 skin biopsies from 34 patients who attended a Renal Transplant Unit in Brisbane over 12 months included 40 squamous cell carcinoma (SCC), 22 solar keratoses, 4 hyperkeratoses, 18 warts and 11 basal cell carcinoma (BCC). Human papillomavirus (HPV) DNA was identified by Southern blot hybridisation using, as individual probes, purified insert DNA from recombinant HPV 1, 2, 3 or 3/10, 4, 5 or 5/8, 7, 11, 16, 18 and 41 under relaxed conditions and characterised by restriction enzyme analysis and Southern blot hybridisation under more stringent conditions. Genomic HPV DNA was characterised in 7 skin biopsies from 4 renal allograft recipients (RARs): HPV 1A in a SCC (20 copies/cell) and a BCC (10 copies/cell) from the one patient, HPV 36 (20 copies/cell) in a SCC, HPV 1A (1000 copies/cell) in a wart and HPV 2B (200-800 copies/cell) in 3 warts from the one patient. Only HPV 1A in the SCC exhibited a significant degree of subtype variation. HPV DNA was identified in another 5 skin biopsies from another 4 RARs: HPV 3A in a wart and a hyperkeratosis, HPV3/10-related DNA in 2 solar keratoses and HPV5/8-related DNA in another (20-50 copies/cell). The incidence of HPV 5 (or 5-related HPVs) in RAR SCC was very low and that of HPV DNA in RAR warts was lower than that recorded elsewhere but this was not due to insensitivity of the assays. There was no evidence for a role for HPV in the aetiology of skin cancer in RARs in south-eastern Queensland but the possibility remains that as yet unidentified HPV types are involved.     

STUDY OF THE PARTNERS OF WOMEN WITH HUMAN PAPILLOMAVIRUS INFECTION

Background. Genital human papillomavirus (HPV) infection is one of the most common sexually transmitted diseases, and it has been identified as a significant risk factor for the development of dysplasia and cancer of the uterine cervix. The possible influence of male HPV lesions on female cervix oncogenesis has not been elucidated so far. In the present study we evaluate the male partners of women with clinical or subclinical HPV infection with particular interest in the clinical features of this infection in both partners. Methods. We examined 81 male partners of women affected with human papillomavirus infections. Condylomata acuminata were searched for by visual inspection. Subclinical lesions were searched by 5 power optical magnification lens after application of 5% acetic acid. Results. In men we observed the following percentage of infection: 67% of the partners of women affected with condylomata acuminata, 46% of the partners of women affected with subclinical lesions (acetic acid positive), and 40% of the partners of women with association of HPV and cervical intraepithelial neoplasia. Conclusions. Our data stress that very often the partners of women with HPV subclinical infection, especially when associated with CIN, do not present lesions, and consequently primary prevention may be very difficult.     

DETECTION WITH THE POLYMERASE CHAIN REACTION OF HUMAN PAPILLOMAVIRUS DNA IN CONDYLOMATA ACUMINATA TREATED WITH CO2 LASER AND MICROWAVE

Background. The recurrence rates of condyloma acuminata are high. The reasons for the relatively high relapse rates with different treatments are unknown. Methods. Twelve specimens of condylomata acuminata of the vulva were excised from 12 patients and divided into three parts. One part was untreated, the second and the third parts were treated with CO₂ laser and microwave, respectively. dna was then extracted from tissue by proteolytic digestion and amplified by the polymerase chain reaction. Dot blots were performed with the use of radiolabeled consensus and human papilloma virus (hpv ) type-specific probes. Results. hpv dna was amplified in 100% of untreated specimens (6-hpv 6; 6-hpv 11), and in 83.3% and 50% of specimens treated with CO₂ laser and microwave, respectively. There was a significant difference in detection between untreated and microwave-treated specimens (χ²= 4.18, P < 0.05). Conclusion. Microwave damages hpv dna more effectively than CO₂ laser.     

A COMPARISON OF IN VIVO AND IN VITRO METHODS OF IDENTIFYING HUMAN EPIDERMAL CELLS IN DNA SYNTHESIS

Summary.— For identifying epidermal cells in DNA synthesis there are 2 rather different methods in use: subcutaneous injection of ³H thymidine in vivo , and short term incubation of skin slices with ³H thymidine in vitro.
A direct comparison of these methods on the skin of the same individuals at the same times shows little difference in the results obtained. If anything, the in vivo method is slightly more sensitive.     

NUCLEAR AND NUCLEOLAR ULTRASTRUCTURE DURING THE LATE STAGES OF NORMAL HUMAN KERATINOCYTE MATURATION

Summary.— Ultrastructural changes of nuclei and nucleoli were studied during the physiological maturation of human keratinocytes. Most nuclear and nucleolar changes were observed in keratinocytes of the upper cell layer of the stratum granulosum, in which these cells are transformed to horny cells. Nucleolar changes were characterized by the segregation of nucleolar ribo-nucleoprotein components, the formation of ring shaped nucleoli and by a reduced quantity of granular components. In addition to the known nuclear ultrastructural changes in keratinocytes of the upper cell layer of the stratum granulosum, an increased number of perichromatin granules was noted in the nuclei of these cells. The number of interchromatin granules was variable.
The nucleolar segregation, formation of ring-shaped nucleoli and the decrease of nucleolar granular components in keratinocytes of the upper cell layer of the stratum granulosum, together with the increase in the number of perichromatin granules, might suggest that the RNA synthesis in these cells is inhibited first in the nucleoli before the cessation of extranucleolar RNA synthesis and nuclear disintegration.     

INCREASE OF ACIDIC NUCLEAR PROTEINS AND DNA SYNTHESIS IN HUMAN LYMPHOCYTES ACTIVATED BY PHYTOHEMAGGLUTININ

Acidic nuclear proteins and histones were extracted at several points in time from human lymphocytes cultured in media containing phytohemagglutinin, and their amounts were measured according to Folin-Lowry's technique. The amount of acidic nuclear proteins continually increased, but the histone level neither increased nor decreased. It is suggested that acidic nuclear proteins might restore histone-inhibited DNA-dependent RNA synthesis in human lymphocytes activated by phytohemagglutinin.     

INHIBITORS AND PROACTIVATORS OF FIBRINOLYSIS IN HUMAN EPIDERMIS

Summary.— Normal human epidermis was homogenized and separated into "heavy mitochondrial", "light mitochondrial", "microsomal" and "soluble" fractions by differential centrifugation in the cold. The homogenate and the subcellular fractions were investigated on fibrin plates for their fibrinolytic activity, their inhibitor activity against urokinase, and proactivator activity.
No direct fibrinolytic activity was demonstrated in the homogenate or the suhcellular fractions, but they showed inhibitory activity against urokinase and strong fibrinolytic activity following streptokinase treatment. Both activities were predominant in the soluble fraction. Weak activity was also found in particulate fractions. The distribution pattern of both activities in subcellular fractions was very similar. The possible existence of a complex of proactivator and inhibitor in the epidermis is discussed.     

银屑病患者血内中性白细胞、单核细胞及淋巴细胞的超微结构观察

本文对15例进行期银屑病患者血内嗜中性白细胞、单核细胞及淋巴细胞通过电镜观察其超微结构的变化、并以10例献血员作对照。发现银屑病患者三种血细胞从细胞形态到细胞内细胞器均有不同程度的异常。经治愈后病人除显性遗传银屑病患者三种血细胞仍保留着治疗前某些特点外,其他基本恢复正常。提示本病不单是皮肤损害,血液内有相应的改变。三种血细胞异常改变是可逆的。     

THE ULTRASTRUCTURE OF THE SKIN OF HUMAN EMBRYOS V. THE HAIR GERM AND PERIFOLLICULAR MESENCHYMAL CELLS HAIR GERM-MESENCHYME INTERACTION

SUMMARY.— Electron microscopic studies on embryonic hair development were performed using the scalp and eyebrow skin of 10 Negro embryos of the menstrual ages of 9 to 15 weeks. New ultramicroscopic findings were: (1) The peripheral cells of the hair germ extended into the mesenchyme pseudopodia, not covered by the basal lamina. (2) Mesenchymal cells crowded beneath the hair germ were connected with desmosome-like junctions. Direct contact of these cells with the basal lamina of the hair germ was seen. (3) A halfdesmosome- like structure was found between the mesenchymal cells and mesenchymal fine filaments.     

银屑病病损皮肤与健康人皮肤cAMP含量测定分析

本文对24例银屑病病损皮肤与12例健康人皮肤进行了cAMP测定,其结果银屑病病损皮肤组织平均含量为0.747pmol/mg,健康人皮肤组织平均含量为0.62pmol/mg。支持银屑病皮肤组织cAMP含量偏低的观点。初步探讨了cAMP对表皮动力学变化的作用。     

ULTRASTRUCTURE OF ADULT EPIDERMAL CELLS IN MONOLAYER CULTURE

SUMMARY.— Epithelial cells from adult guinea-pig and human skin were grown in monolayer culture and studied by phase contrast and electron microscopy. Significant differences were found between cells in vivo and in vitro .
Keratinocytes show a selective dedifferentiation. They retain the ability to make desmosomes, but interdigitation is reduced and the system of tono-filaments is disorganized. Mitochondria become long and even branched-while microtubules become prominent, as do lysosomes and fat droplets. Ribosomes often become arranged in polyribosomes. and channels appear in the nucleus. Melanocytes showed similar changes, and 1 produced a cilium.
Such changes should be taken into consideration when cells in dispersed culture are used experimentally.