Changes in surfactant protein A and D in ovine ovaries related to follicle development.

Surfactant protein A (SP-A) and Surfactant protein D (SP-D) glycoproteins play a crucial role in maintaining lung homeostasis and lung host defense. Interestingly, these proteins are also expressed in extra-pulmonary tissues, including the female genital tract. The ovarian tissue, where SP-A and SP-D expression increases with follicular development, may serve as the primary site of defense for this tissue. However, their functions in these tissues are not well understood and are currently an active area of research. Therefore, the objective of this study is to investigate the expression of SP-A and SP-D in the ovine ovary throughout the ovarian cycle using immunohistochemistry by semiquantitative intensity classification and Western blotting techniques. These findings revealed the presence of SP-A and SP-D in various compartments of the ovary, such as the follicular epithelium, granulosa cells, cumulus cells, theca cells, oocyte I, follicular fluid, and luteal cells of Graafian follicles, excluding the corpus albicans. SP-A and SP-D likely act as a first line of defense against potential pathogens that infiltrate the ovaries. Further investigation of the differential expression of SP-A and SP-D proteins in ovarian follicles will provide a basis for understanding their interactions with key proteins involved in oogenesis.

Evidence of surfactant protein A and D expression decrement and their localizations in human prostate adenocarcinomas.

AIM: Surfactant proteins (SP-A and SP-D) were originally described in the lung; however, they are also present in the prostate. Purpose of this study, therefore, was to determine how surfactant proteins are altered in prostate adenocarcinomas (PCa) and find out any connection exists between their expressions and their staining patterns, prostate-specific antigen (PSA) values, Gleason score, age, tumor volume and tumor, node, metastases (TNM) clinical stage. METHODS: Thirty-five tissue samples were obtained during radical prostatectomy. All specimens were classified to three groups based on the Gleason score <7, 7 and Gleason score >7. Surfactant proteins' expressions were tested by immunohistochemical and Western blotting methods. RESULTS: Immunoreactivity was detected in the cytoplasm from both basal cells and secretory epithelial cells in malignant and non-malignant areas. About 80% of the malignant basal cells were characterized as either weak or strong while non-malignant epithelial cells demonstrated strong immunoreactivity for SP-A. Also malignant (81.8%) and non-malignant cells (90.6%) were characterized as either weak or strong for SP-D. Decrement of SP-A and SP-D immunostaining tended to associate with an increasing Gleason score (p > 0.05, p < 0.05), tumor volume (p < 0.05, p > 0.05) and age (p > 0.05, p > 0.05). There was a strong positive correlation between Gleason score and tumor volume (p < 0.01). Also, either none or weak SP-A and SP-D immunoreactivity was observed specimens with Gleason score 7 or higher. SP-A and SP-D reacted with 34 kDa (SP-A) and 43 kDa (SP-D) immunoreactive single bands were decreased in tumor tissues. CONCLUSIONS: The development of prostate cancer may be related to decreased level of surfactant protein A and D.

Surfactant proteins in inflammatory skin diseases: controlled study.

Surfactant proteins (SP) have recently been reported to be expressed in human skin tissue. SP is thought to play an essential role in the firstline defense of skin. In this study, we aimed to investigate if the SP may play a role in inflammatory skin diseases. Seven volunteers with psoriasis (n = 3), atopic dermatitis (n = 2), lichen planus (n = 1) and Behcet's disease (n = 1) participated in the study. Biopsies from each lesion and adjacent (approximately 2 cm distant) normal-appearing skin in patients were performed. Expression and localization of the SP-A, -B, -C, and -D in fresh tissues were studied by an immunohistochemical technique. In all patients, there was a weak cytoplasmic staining with SP-A and SP-D and nuclear staining with SP-B and SP-C in the epidermis of normal-appearing skin samples. However, epidermal staining with SP was observed to be stronger in all lesional samples. In addition, there was a prominent staining in inflammatory cells infiltrating dermis. This expression represents a previously unknown immunologic response in the inflammatory skin diseases and may represent an important step in the pathogenesis of these disorders.

Determination of the expression of fish antifreeze protein (AFP) in 7th generation transgenic mice tissues and serum.

In this study, the presence of antifreeze protein (AFP) gene expression through successive generations in transgenic mice carrying the chimeric gene construct of the coding sequence for the AFP protein from ocean pout was investigated. AFP transgenic hemizygote mice were used for AFP gene expression. AFP genome expressions in transgenic mice were analyzed by Western blotting, and tissue location of AFP protein was shown by immunohistochemical and immunofluorescence techniques. Seventh transgenic mice from the established founders demonstrated the expression of AFP in organs such as the skin, oviduct, lung, kidney and liver tissues and serum except for the heart. Our results demonstrate successful expression of AFP gene products in several tissues and serum of transgenic mice, the association of in vivo expressed AFP protein, for the first time. These results indicate that the coding sequence for the AFP protein gene (ocean pout type III AFP gene) could be integrated and stably transcribed and expressed in the 7th generation of transgenic mice. In conclusion transgenic mouse lines would be a good model for the cryostudy of AFP and for the determination of AFP roles in several organs and tissues.

Presence and subcellular localizations of surfactant proteins A and D in human spermatozoa.

OBJECTIVE: To investigate the presence of surfactant protein-A (SP-A); molecular weight 34 kDa and surfactant protein-D (SP-D); and molecular weight 43 kDa in human spermatozoa. DESIGN: Prospective, research study. SETTING: Two universities in Turkey. PATIENT(S): Semen specimens (n = 10) were obtained from normozoospermic donors. INTERVENTION(S): Human sperm were exposed to an anti-human SP-A polyclonal antibody, and monoclonal antibody, to human SP-D protein. MAIN OUTCOME MEASURE(S): Presence of SP-A and SP-D proteins in human beings. RESULT(S): Indirect immunofluorescence assays of human sperm indicated the presence of SP-A in the mid piece, the tail, and sometimes at the equatorial region of spermatozoa. A brilliant green light detected SP-D in the tails and acrosome of some sperm. The anti-SP-A antibody detected a single band corresponding to the molecular weight values of 34 kDa in spermatozoa, whereas no band was observed in the negative control. The anti-SP-D antibody showed the expected band at 43 kDa in spermatozoa. CONCLUSION(S): This is the first report and a novel finding of the presence of surfactant glycoproteins on human spermatozoa.

Surfactant protein expression in human skin: evidence and implications.

The presence of surfactant proteins (SPs), critical to local barrier and defense functions and usually associated with the lung, was revealed in adult and fetal human skin complementary deoxyribonucleic acid, in skin samples from three adult female donors and also in cultured fibroblasts, keratinocytes, and melanocytes. Using reverse transcription-PCR, SP-A, SP-B, SP-C, and SP-D messenger ribonucleic acid expression was detected to varying extents in the different skin sources. The stronger expression of SP-C in fetal skin, compared to adult skin, suggested that the role of this protein alters with age. Immunohistochemical studies showed variable distribution of SPs in human epidermis and dermis, confirming that these proteins are indeed translated and expressed in skin tissue. In vitro studies showed that the surface tension of SP-deficient artificial sebum is (a) lowered by skin-extracted SP-B and (b) further reduced to a level comparable to normal sebum by the additional presence of skin-extracted SP-A and SP-D, consistent with their surface tension-lowering capabilities in lung. The possible roles of SPs in skin, based on their known functions in the lung are discussed. However, their potential as therapeutic targets or diagnostic markers of skin disease remains to be elucidated.

Surfactant proteins A and D in the genital tract of mares.

The presence of surface-active material in the lung alveolus has been known for several decades as being essential for normal lung function. Surfactant is essential for reducing the surface tension at the alveolar air-liquid interface. Pulmonary surfactant is composed of 90% lipids and 10% proteins. There are four non-serum proteins surfactant protein-A (SP-A), surfactant protein-B (SP-B), surfactant protein-C (SP-C) and surfactant protein-D (SP-D) named in chronologic order of discovery. Lung SP-A and SP-D belong to a family of collagen-containing C-type lectin family called collectins. The host defence and controlling inflammatory processes of the lung are the major functions of SP-A and SP-D. SP-A and SP-D were originally demonstrated in alveolar type II cells, but recent studies have shown extrapulmonary expression of SP-A and SP-D indicating systemic roles of these proteins. Present study describes the presence of SP-A and SP-D in the mare genital tract, vulva, vagina, ovarium, uterus and tuba uterina using immunohistochemistry and Western blotting. The aim of this study was to characterize surfactant proteins in terms of: (i) whether surfactant proteins were present in the various structures of the mare genital system, (ii) if so, identifying and locating the surfactant proteins and finally (iii) determining the differences from those previously characterized for the lung. Although beyond the scope of this report, it is recognized that there are also some potential implications for better defining the reproductive defence mechanisms in mare. Therefore, genital system organs and tissues from mares were examined. We were able to show that proteins reactive with surfactant-specific antibodies were present in the mare genital tract. Thus, surfactant proteins are present not in just lamellar bodies associated with lung, but also genital system of mare.

Surfactant protein A and D in the reproductive tract of stallion.

The presence of surface-active material in the lung alveolus has been known for several decades as being essential for normal lung function. The host defense and controlling inflammatory processes of the lung are the major functions of SP-A and SP-D. SP-A and SP-D were originally demonstrated in alveolar type II cells, but recent studies have shown extrapulmonary expression of SP-A and SP-D indicating systemic roles of these proteins. Present study describes the presence of SP-A and SP-D in the stallion genital tract, prepuce, prostate, testis, and seminal vesicle using Western blotting and immunohistochemistry. This paper presents the first evidence for the existence of SP-A and SP-D glycoproteins in the stallion genital tract. We examined genital system organs and tissues from stallion and were able to show that surfactant protein A and D reactive with surfactant-specific antibodies were present in the stallion genital tract tissues and organs. On the basis of results, it can be postulated that surfactant proteins in the stallion reproductive tract contribute to the immune surveillance and to active barrier defense mechanism.

Increased expression of surfactant protein A and D in rheumatoid arthritic synovial fluid (RASF).

AIM: To determine the concentrations of surfactant protein (SP)-A and SP-D in synovial fluid samples from patients with rheumatoid arthritis and healthy controls and correlate them with rheumatoid factor, C-reactive protein (CRP), immunoglobulin (Ig) A, IgM, and IgG, total protein, and total lipid content. METHODS: The concentrations of SP-A, SP-D, CRP, IgA, IgM, IgG, and rheumatoid factor were measured in the synovial fluid of 7 patients with rheumatoid arthritis and 3 samples of healthy synovial fluid obtained at autopsy. The bands obtained by specific antibodies in Western blotting to determine the surfactant protein concentration were analyzed densitometrically and synovial fluid total phospholipids and protein content were analyzed spectrophotometrically. RESULTS: Patients with rheumatoid arthritis had increased concentrations of proteins and lipids in the synovial fluid, which correlated with 3.5-fold increase in SP-A and 6.1-fold increase in SP-D concentrations. Total protein content in synovial fluid samples from patients with rheumatoid arthritis showed a 2.1-fold increase and phospholipid content showed 7.0-fold increase in comparison with samples of healthy synovial fluid. Rheumatoid factor, CRP, IgA, IgM, and IgG concentrations in synovial fluid samples from patients with rheumatoid arthritis were 40-2660 KIU/L, 4-35 mg/L, 0.10-2.70 g/L, 0.50-1.90 g/L, and 5.3-15.4 g/L respectively. CONCLUSION: SP-A and SP-D were present and expressed in various degrees in the synovial fluid of patients with rheumatoid arthritis. SP-A and SP-D may play role in the initiation of immune system and joint inflammation, and may be an integral component of synovial fluid and provide insights for in innate and adaptive immunity within the joint.

Detection of surfactant protein A (SP-A) and surfactant protein D (SP-D) in equine synovial fluid with immunoblotting.

Once considered unique to the lung, surfactant proteins have been clearly identified in the intestine and peritoneum and are suggested to exist in several other organs. In the lung, surfactant proteins assist in the formation of a monolayer of surface-active phospholipid at the liquid-air interface of the alveolar lining, reducing the surface tension at this surface. In contrast, surface-active phospholipid adsorbed to articular surfaces has been identified as the load-bearing boundary lubricant of the joint. This raises the question of whether surfactant proteins in synovial fluid (SF) are required for the formation of the adsorbed layer in normal joints. Proteins from small volumes of equine SF were resolved by 1- and 2-dimensional polyacrylamide gel electrophoresis and detected by Western blotting to investigate the presence of surfactant proteins. The study showed that surfactant proteins A and D (SP-A and SP-D) are present in the SF of normal horses. We suggest that, like surface-active phospholipid, SP-A and SP-D play a significant role in the functioning of joints. Next will be clarification of the roles of surfactant proteins as disease markers in a variety of joint diseases, such as degenerative joint disease and inflammatory problems.

Immunodetection of surfactant proteins in human organ of Corti, Eustachian tube and kidney.

The presence of surfactant proteins was investigated in the human organ of Corti, Eustachian tube and kidney tissues. It has previously been shown that lamellar bodies are present in hairy cells of organ of Corti, in the cytoplasm of secretory and lumen of tubal glands of Eustachian tube and kidney renal basement membrane. No evidence for the presence of surfactant proteins in the organ of Corti and kidney has been presented until now. The aim of this study was to find out if surfactant proteins were expressed in other epithelia such as organ of Corti, Eustachian tube and kidney. Surfactant proteins were identified using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. On one-dimensional Western blots, bands for surfactant protein A in human Eustachian tube (SP-A, 34 kDa) and in kidney extracts, and for surfactant protein D (SP-D, 43 kDa) in Eustachian tube and in kidney extracts (SP-D, 86 kDa), and for surfactant protein B (SP-B, 8 kDa) in human Eustachian tube and organ of Corti extracts were detected. Bands corresponded to monomeric forms of lung surfactant proteins. These results indicate the presence of SP-A and SP-D in kidney epithelium, SP-A, SP-B and SP-D in Eustachian tube and SP-B in the organ of Corti.