OBJECTIVE: Multiple morphological abnormalities of the sperm flagella (MMAF) are a severe form of sperm defect causing male infertility. Previous studies identified the variants in the CFAP69 gene as a MMAF-associated factor, but few cases have been reported. This study was performed to identify additional variants in CFAP69 and describe the semen characteristics and outcomes of assisted reproductive technology (ART) in CFAP69-affected couples.
METHODS: Genetic testing with next-generation sequencing (NGS) panel of 22 MMAF-associated genes and Sanger sequencing was performed in a cohort of 35 infertile males with MMAF to identify pathogenic variants. Morphological, ultrastructural, and immunostaining analyses were performed to investigate the characteristics of probands\' spermatozoa. ART with intracytoplasmic sperm injection (ICSI) was carried out for the affected couples to get their own progenies.
RESULTS: We identified a novel frameshift variant in CFAP69 (c.2061dup, p. Pro688Thrfs*5) from a MMAF-affected infertile male with low sperm motility and malformed morphology of sperm. Furthermore, transmission electron microscopy and immunofluorescence staining revealed that the variant induced the aberrant ultrastructure and reduction of CFAP69 expression in the proband\'s spermatozoa. Moreover, the partner of the proband birthed a healthy girl through ICSI.
CONCLUSIONS: This study expanded the variant spectrum of CFAP69 and described the good outcome of ART treatment with ICSI, which is beneficial to the molecular diagnosis, genetic counseling, and treatment of infertile males with MMAF in the future.

The protein encoded by dynein axonemal heavy chain 1 (DNAH1) is a part of dynein, which regulates the function of cilia and sperm flagella. The mutant of DNAH1 causes the deletion of inner dynein arm 3 in the flagellum, leading to multiple morphological abnormalities of the sperm flagella (MMAF) and severe asthenozoospermia. However, instead of asthenozoospermia and MMAF, the result caused by the mutation of DNAH1 remains unknown. Here we report a male infertility patient with severe asthenozoospermia and teratozoospermia. We found two heterozygous mutations in DNAH1 (c.6912C>A and c.7076G>T) and which were reported to be associated with MMAF for the first time. We next collected and analyzed 65 cases of DNAH1 mutation and found that the proportion of short flagella is the largest, while the bent flagella account for the smallest, and the incidence of head deformity is not high in the sperm of these patients. Finally, we also analyzed 31 DNAH1 mutation patients who were treated with intracytoplasmic sperm injection (ICSI) and achieved beneficial outcomes. We hope our research will be helpful in the diagnosis and treatment of male infertility caused by DNAH1 mutation.

OBJECTIVE: To determine the meiotic segregation in large-headed, multiple-tailed spermatozoa.
METHODS: Analysis of sperm nuclei by fluorescence in situ hybridization (FISH).
METHODS: University hospital.
METHODS: A 34-year-old man with 100% morphologically abnormal spermatozoa.
METHODS: Dual-color FISH for chromosomes 13 and 21 and triple-color FISH for chromosomes X, Y, and 18 were performed.
METHODS: Aneuploidy rates.
RESULTS: More than 99% of the spermatozoa had abnormal content for chromosomes X, Y, 13, 18, and 21. Diploidy, triploidy, and tetraploidy rates were found to be 18.42%, 6.14%, and 33.99% in triple-color FISH and to be 16.09%, 16.28%, and 38.95% in dual-color FISH.
CONCLUSIONS: Our results and those from other investigators show that large-headed, multiple-tailed spermatozoa are associated with a high rate of polyploidy and aneuploidy. Intracytoplasmic sperm injection should not be recommended to those patients, not only because of its low success rate but also because of its high genetic risk.

The identification of human sperm hyperactivated motility has potential importance in sperm function tests, as well as in quality control assays and in reproductive toxicology investigations. However, relatively little is known about this phenomenon and the variety of definitions used for hyperactivation has led to a great deal of confusion as to its occurrence and physiological relevance. This presentation is a critical review of a number of aspects of hyperactivated motility, including its identification and potential role(s) in mammalian fertilization. The initial sections of the review consider the mechanisms involved in the development and maintenance of mammalian sperm motility, and the structural and functional changes in spermatozoa which occur during transport through the female reproductive tract. The methods available for the quantification of aspects of sperm movement are also discussed, with an historical overview of sperm movement analysis.

The cardiac glycoside, ouabain, exerts its influence on spermatozoa by binding to and inhibiting Mg2+-activated Na+, K+-dependent ATPase that is located in the midpiece-tail membranes. Ouabain decreased intracellular potassium, increased intracellular sodium, and produced a biphasic and time-dependent effect on motility--stimulation at low concentrations and inhibition at high concentrations. The motility depression consisted of decreases in numbers of motile cells, percent progressive motility, beat frequency, and amplitude. Species differences and maturational age of the sperm cells were reflected in the degree of the ouabain effect and also the distribution of the ouabain-sensitive enzyme. The presence of this enzyme in spermatozoan membranes contributes significantly to regulation of sperm cell function through modulation of cationic fluxes which in \"conventional\" cell types influence their excitability.

A case of immotile cilia syndrome (ICS) is presented. A 34-year-old male, who had suffered from recurrent bronchitis, sinusitis and otitis media since early childhood, was admitted to Keio University Hospital complaining of productive cough and infertility. A saccharin test showed prolonged nasal clearance time, and semen analysis revealed immotile sperm. By electron microscopic observation of cilia of the nasal mucous epithelium and the sperm, inner and outer dynein arm defect, abnormal microtubular arrangement and compound cilia were detected and he was diagnosed as ICS. Thirty eight Japanese cases of ICS collected from the literature were analyzed concerning clinical manifestations, airway clearance, family history and ciliary ultrastructural abnormalities (Table 2, 3). Recurrent bronchitis, male infertility, chronic sinusitis, otitis and decrease in airway clearance were very common in these patients. Female infertility was more common than anticipated. The prevalence rate of situs inversus was more than 50%, probably due to more attention having been paid to Kartagener\'s syndrome in Japan. Recent studies show that the cilia of this syndrome is not always immotile but characterized by hypomotility or asynchrony, and have attempted to determine the relationship between each ciliary ultrastructural abnormality and motility pattern. It is necessary that more patients including incomplete and mild cases should be investigated.