ABSTRACT
Dihydrotestosterone (DHT) binding to androgen receptors (AR) in hair follicles is commonly accepted as the first step leading to the miniaturizing of follicles associated with androgenetic alopecia (AGA). Testosterone is converted to DHT by the enzyme 5alpha-reductase. Finasateride a 5alpha-reducase inhibitor blocks the production of DHT and is currently used to treat AGA. The inhibition is not complete but a reduction of DHT systemically and in the scalp is accomplished. Ketoconazole has been clinically shown to be effective in the treatment of AGA. In this paper, evidence is presented to support the hypothesis that ketoconazole 2% shampoo has a local disruption of the DHT pathway. It is proposed that using ketoconazole 2% shampoo as an adjunct to finasteride treatment could lead to a more complete inhibition of DHT and thus better treat AGA.
Subject(s)
Alopecia/drug therapy , Evidence-Based Medicine/methods , Finasteride/administration & dosage , Ketoconazole/administration & dosage , Administration, Oral , Administration, Topical , Antifungal Agents/administration & dosage , Chemotherapy, Adjuvant/methods , Clinical Trials as Topic , Drug Therapy, Combination , Enzyme Inhibitors/administration & dosage , Hair Preparations , Humans , Male , Treatment OutcomeABSTRACT
Spermiophages were isolated from turkey semen using Percoll gradient centrifugation, cultured in Roswell Park Memorial Institute 1640 medium at room temperature, and characterized by transmission electron microscopy. After 1 to 2 h, the cells enlarged and developed numerous motile mitochondria. Over time, the mitochondria appeared to increase in number and were released into the extracellular medium. Few mitochondria were observed in spermiophages from fresh semen. However, there was an apparent increase in the number and size of mitochondria after Percoll isolation, which was more pronounced in cultured spermiophages. Over a period of 3 h in culture, many spermiophages became engorged with mitochondria, which subsequently appeared to be released as blebs pinched off from the surface. The release of mitochondria resulted in spermiophages with large, empty vacuoles, although their remaining cytoplasm was engorged with mitochondria. Many free mitochondria were present in the medium. The results of the current research suggest that isolating and culturing turkey spermiophages elicit mitochondrial biogenesis, which proceeds unabated until the cells are engorged with and release numerous mitochondria. This may be due to conditions under which the spermiophages were cultured or to nonhistocompatibility of these cells in pooled semen.
Subject(s)
Macrophages/ultrastructure , Semen/cytology , Turkeys , Animals , Male , Microscopy, Electron , Mitochondria/ultrastructureABSTRACT
A procedure was developed to rapidly isolate functional, intact mitochondria from turkey spermatozoa. Semen was collected from turkeys, pooled, and centrifuged to remove spermiophages and other cells. The sperm cells were then mechanically disrupted with a Dounce homogenizer, sonicated, and centrifuged using a discontinuous Percoll gradient. Electron microscopy revealed morphologically intact mitochondria. The isolated mitochondria exhibited cytochrome oxidase activity, oxygen consumption, and were stained by rhodamine 123, a fluorescent stain specific for functional mitochondria in eukaryotic cells. Mitochondrial DNA (mtDNA) was isolated and purified, and the genome was determined to be 16.457 +/- 0.07 kbp. Restriction fragment patterns were identified using the endonucleases EcoR1, HindIII, and BamH1. Mitochondrial DNA was also purified from turkey liver and testis, and no differences in the restriction enzyme patterns were found between somatic and germ cell mtDNA. It is concluded that mitochondria can be isolated from spermatozoa for metabolic or genetic study.
