Presence of antisperm antibodies in the serum of Sardinian adult people. II. Study on a selected infertile population in north Sardinia.

Among many possible causes of infertility an immunological factor must be considered, in particular the presence of antibodies to spermatozoa in the serum. In previous researches we confirmed the presence of antisperm antibodies in the serum of healthy men of North Sardinia in percentage equal to 2.4% of the considered population. The aim of the present study is to evaluate the correlation between the presence of antisperm antibodies and infertility. The presence of antispermatozoal antibodies has been evaluated in the serum of 124 men (aged 25-50) affected by unexplained infertility utilizing the indirect immunofluorescence and ELISA methods. The presence of antisperm antibodies has been found equal to 19.3% of the tested sera. These results show that the incidence of antisperm antibodies is higher in the serum of male partners from infertile couples than in healthy subjects (P < 0.01), also in a relatively segregated and pure population like the one studied.     

La stérilité d'origine immunologique.

Antispermatozoal antibodies in the woman, the man, or both, may be the cause of nonorganic infertility in a small number of couples. The antibodies are most often found in the serum or in the genital fluids. Diagnosis is based on the postcoital test and on various laboratory tests to assess sperm-cervical mucous compatibility and to detect antispermatozoal antibodies. In women the strongest relation between results of laboratory tests and infertility is between the presence of serum antispermatozoal antibodies and infertility of more than 3 years'' duration. In men there is a relation between high levels of these antibodies in the semen and infertility. Various treatments have been proposed, but further prospective studies are needed. However, promising results have been reported with homologous intrauterine insemination and with immunosuppression in men.     

Body weight and risk of myocardial infarction and death in the adult population of eastern Finland.

Body mass index (weight (kg) divided by height squared (m2] and its association with the risk of myocardial infarction and death from all causes were studied prospectively in a randomly selected population sample in eastern Finland aged 30-59 at outset in 1972. The study population consisted of 3786 men and 4120 women. The participation rate in the survey in 1972 was over 90%. All deaths and admissions to hospital in the sample were obtained from the National Death Certificate and Hospital Discharge Registers. During the seven years of follow up until 1978, 170 men and 52 women had acute myocardial infarction, and during the nine years up to 1980, 223 men and 92 women died. Independent of age, men with a body mass index of 28.5 or more had a significantly higher incidence of acute myocardial infarction. This effect was also independent of smoking but not independent of biological coronary risk factors--that is, serum cholesterol concentration and blood pressure. In the analysis stratified for smoking in men the body mass index total mortality curve was J shaped among non-smokers, whereas smoking entirely outweighed body mass index as a predictor of death. Body mass index did not contribute significantly to the risk of either acute myocardial infarction or death in women. It is concluded that a body mass index of around 29.0-31.0 or more is not only a marker for coronary risk factors but is also a predictor of acute myocardial infarction in men.     

Reference values of thyroid volume in a healthy, non-iodine-deficient Spanish population.

Our aim was to assess reference values of thyroid volume by ultrasonography in healthy adult subjects. We conducted an epidemiological cross-sectional study where 880 subjects were randomly selected from the town census of L'Hospitalet de Llobregat after being invited to participate in our study directly by mail and phone call. We made a clinical history of each subject and determined serum thyrotropin, antiperoxidase antibodies, urinary iodine excretion and thyroid volume by ultrasonography. Subjects with thyroid disease were excluded. We finally studied 268 representative subjects. The reference thyroid volume was median 7.31 ml, mean 8.22 ml (Confidence Interval: 7.75 - 8.69 ml). In men: median 9.19 ml, mean 9.87 ml (CI: 9.09 - 10.65 ml); in women: median 6.19 ml, mean 6.57 ml (CI: 6.22 - 9.92 ml) (p < 0.0001). We grouped the subjects into decades, and found that thyroid volume was different (p = 0.0034) in males because the younger group had lower volume. We did not find any differences among age groups in women. The mean of the urinary iodine excretion was 154.23 microg/l. We have determined reference values of thyroid volume measured by ultrasonography in our iodine non-deficient population and prepared tables that distribute thyroid volume by sex and age.     

A novel approach to the generation of antibodies against phylogenetically preserved sperm antigens

Conventional methods for immunization of laboratory animals against human spermatozoa proved not to be efficient enough to identify phylogenetically conserved sperm-specific antigens. A combination of vasectomy and subcutaneous administration of autologous testis homogenates was tested in 5 New-Zealand rabbits, and in 7 Long-Evans and 8 Spraque-Dawley rats in an attempt to induce an autoimmune response against such antigens. This experimental procedure resulted in a generation of sperm autoantibodies cross-reactive with human, rabbit and rat spermatozoa, as demonstrated by sperm-agglutination, ELISA and flow cytometry (FCM). No specific binding to human seminal plasma was detected by ELISA, indicating that intrinsic sperm membrane antigens rather than sperm-coating antigens were involved in establishing cross-reactivity with human spermatozoa. This suggestion was confirmed by the finding that rabbit autoantisera reacted more strongly against epididymal than against ejaculated human spermatozoa as shown by FCM. Humoral antispermatozoal response correlated well with impaired spermatogenesis in rabbits. The autoimmunized rats revealed severe alterations in reproductive tissues, including testicular and epididymal sperm granulomas; however, they showed a lower incidence of circulating antibodies. The results indicate that the established experimental model in rabbits can be further used to identify and characterize evolutionary preserved intrinsic sperm membrane autoantigens, which are desirable candidates for contraceptive vaccine development.     

Failure of the expression of phospholipid hydroperoxide glutathione peroxidase in the spermatozoa of human infertile males

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) was intensely expressed in mitochondria in the midpiece of human spermatozoa by immunostaining with anti-PHGPx monoclonal antibodies. The PHGPx not only reduced phospholipid hydroperoxide but also scavenged hydrogen peroxide in human spermatozoa. We found a dramatic decrease in the level of expression of PHGPx in the spermatozoa of some infertile males by immunoblotting with anti-PHGPx monoclonal antibodies. These individuals accounted for about 10% of the group of 73 infertile males that we examined. All seven patients with PHGPx-defective spermatozoa were classified as suffering from oligoasthenozoospermia, a defect in which both the number and the motility of spermatozoa are significantly below normal. Males with PHGPx-defective spermatozoa accounted for 26% of the 27 infertile males with oligoasthenozoospermia. No defects in expression of PHGPx in spermatozoa were observed in 31 fertile volunteers. After a 3-h incubation, the relative number of motile spermatozoa with low-level expression of PHGPx was significantly lower than that of spermatozoa with normal expression of PHGPx. The PHGPx-defective spermatozoa failed to incorporate rhodamine 123, revealing a loss of mitochondrial membrane potential. Ultrastructual analysis of mitochondria by electron microscopy demonstrated that the morphology of mitochondria in PHGPx-defective spermatozoa was abnormal. The results suggest that failure of the expression of mitochondrial PHGPx in spermatozoa might be one of the causes of oligoasthenozoospermia in infertile men.     

Spouse selection by health status and physical traits. Sardinia, 1856–1925

Military medical information and data from civil registers of death and marriage have been used to study the role of physical characteristics and health conditions in explaining access to marriage for the male population of Alghero, a small city located in Sardinia Island (Italy), at the turn of 19th century. Literature data about contemporary populations have already demonstrated the influence of somatic traits in the mate choice. The results presented here show that men with low height and poor health status at the age of 20 were negatively selected for marriage. This holds true also in a society where families often arranged marriages for their children. This pattern of male selection on marriage was found to be particularly marked among the richest and wealthiest SES groups. Our hypothesis is that this social group carefully selected for marriage those individuals who were apparently healthier and therefore more likely to guarantee good health status and better life conditions to offspring. In evolutionary terms, the mate choice component of sexual selection suggests that the height of prospective partners could be claimed as one of the determinants, along with other environmental causes, of the observed higher stature of men belonging to the wealthiest social strata of the Alghero population. Am J Phys Anthropol, 2010. © 2009 Wiley‐Liss, Inc.     

Effect of coculturing spermatozoa with oviductal cells on the incidence of polyspermy in pig in vitro fertilization

It is known that oviductal cells play an important role in fertilization in vivo. We were interested in the effect of those cells on spermatozoa and their influence on the incidence of polyspermy in pig in vitro fertilization (IVF) when cocultured with spermatozoa. Oviductal cells are believed to select a highly fecund population of spermatozoa. By coculturing spermatozoa with oviductal cells it is possible to reduce the number of spermatozoa confronting the eggs at fertilization, reducing the incidence of polyspermy. In our study, spermatozoa were cocultured with oviductal cells for 30 min so that they could bind to the oviductal cells. Both bound and unbound spermatozoa were used for fertilization. The results show that the spermatozoa that were bound to the oviductal cells were capable of fertilizing the eggs, but the nonbound spermatozoa had a reduced penetration incidence. With the bound sperm, polyspermy incidence decreased and the two-pronuclei proportion increased. We also found that with time the spermatozoa released from the cells had better motility than those that did not bind. Therefore, it is our belief that oviductal cells can be used for porcine IVF resulting in a lower polyspermy incidence and higher pronuclei incidence. © 1995 Wiley-Liss, Inc.     

Chromosome 21 disomy in the spermatozoa of the fathers of children with trisomy 21, in a population with a high prevalence of Down syndrome: increased incidence in cases of paternal origin.

Between April 1991 and December 1994, epidemiological studies detected a population with a high prevalence of Down syndrome in El Vallès, Spain. Parallel double studies were carried out to determine the parental and the meiotic origins of the trisomy 21, by use of DNA polymorphisms, and to establish the incidence of disomy 21 in the spermatozoa of the fathers of affected children, by use of multicolor FISH. Results show that the overall incidence of chromosome 21 disomy in the fathers of affected children was not significantly different from that in the control population (0.31% vs. 0.37%). However, analysis of individual data demonstrates that two cases (DP-4 and DP-5) with significant increases of disomy 21 (0. 75% and 0.78% vs. 0.37%) correspond to the fathers of the two individuals with Down syndrome of paternal origin. DP-5 also had a significant increase of sex-chromosome disomies (0.69% vs. 0.37%) and of diploid spermatozoa (1.13% vs. 0.24%).     

Types of structural chromosome aberrations and their incidences in human spermatozoa X-irradiated in vitro

We studied the effects of in vitro X-irradiation on human sperm chromosomes, using our interspecific in vitro fertilization system between human spermatozoa and zona-free hamster oocytes. 28 semen samples from 5 healthy men were exposed to 0.23, 0.45, 0.91 and 1.82 Gy of X-rays. Totals of 2098 and 2862 spermatozoa were karyotyped in the control and the irradiated groups, respectively. The incidence of spermatozoa with X-ray-induced structural chromosome aberrations (Y) increased linearly with increasing dosage (D), being best expressed by the equation, Y = 0.08 + 34.52 D. The incidence of breakage-type aberrations was more than 9 times higher than that of exchange-type aberrations. Both of them showed linear dose-dependent increases, which were expressed by the regression lines, Y = -0.014 + 0.478 D and Y = -0.010 + 0.057 D, respectively. The incidence of chromosome-type aberrations was about 6 times higher than that of chromatid-type aberrations. Their dose-dependent increases were expressed by the regression lines, Y = -0.015 + 0.462 D and Y = -0.006 + 0.079 D, respectively. These results are discussed in relation to the previous data obtained with gamma-rays. The repair mechanism of X-ray-induced sperm DNA lesions is also discussed.     

Increased Frequency of Aneuploidy in Long-Lived Spermatozoa

Aneuploidy commonly causes spontaneous abortions, stillbirths, and aneuploid births in humans. Notably, the majority of sex chromosome aneuploidies in live births have a paternal origin. An increased frequency of aneuploidy is also associated with male infertility. However, the dynamics and behavior of aneuploid spermatozoa during fertilization in humans have not been studied in detail. Therefore, we compared the frequency of aneuploidy and euploidy in live spermatozoa from normozoospermic men over a 3-day period. To assess the dynamics and behavior of aneuploid spermatozoa, we simultaneously evaluated sperm viability using the hypo-osmotic swelling test and sperm aneuploidy using fluorescence in situ hybridization. Whereas the frequency of viable euploid spermatozoa significantly decreased over 3 days, the frequency of viable spermatozoa with aneuploidy interestingly showed a time-dependent increase. In addition, spermatozoa with abnormal sex chromosomes survived longer. To compared with spermatozoa with other swelling patterns, those with tail-tip swelling patterns had a lower frequency of aneuploidy at all time points. This study revealed the novel finding that the frequency of aneuploid spermatozoa with fertilization capability significantly increased compared to that of euploid spermatozoa over 3 days, suggesting that aneuploid spermatozoa can survive longer than euploid spermatozoa and have a greater chance of fertilizing oocytes.     

Restriction polymorphism in patients with lipid metabolism disorders and ischemic heart disease

Using the RELP analysis we studied the frequency of X2 allele of apoB gene in three groups of patients: 1) men at the age of 20-59 with lipid metabolism disorders revealed in population inspection of Oktyabrsky district in Moscow; 2) men with ischaemic heart disease and 3) healthy men. It was established that in individuals suffering from type IIa hyperlipidemia the frequency of X2 allele was significantly higher than in healthy donors from Moscow population. Homozygotes for X2 allele of XbaI RELP had 7-9% higher serum cholesterol levels, than homozygotes for X1 allele. The study suggests the X2 allele of the apoB gene to be associated with the development of high plasma cholesterol level. No significant difference in X2 allele frequencies was found between patients with ischaemic heart disease and healthy donors. There was also no association found between cholesterol and triglyceride levels and the presence of X2 allele in this group of patients.     

Expression of gp20, a human sperm antigen of epididymal origin, is reduced in spermatozoa from subfertile men

gp20, a sialylglycoprotein of human sperm homologous to CD52, is present everywhere on the surface of the freshly ejaculated sperm but is prevalently localized in the equatorial region of the head of capacitated sperm. In the present study, we confirmed this feature on large scale and correlated equatorial exposure of the antigen to the presence of serum albumin (SA) in the capacitation medium. Furthermore, we analyzed the relationship between the presence of the antigen and its equatorial exposure after capacitation and fertility, by comparing immunostaining for gp20 in the motile fraction of spermatozoa from fertile and subfertile men. A significantly higher percentage of nonimmunostained spermatozoa before capacitation (38.5% +/- 23 vs. 12% +/- 7, P < 0.0001) and a lower increase in the percentage of sperm with equatorial localization after capacitation (19.3% +/- 25 vs. 34.6% +/- 22, P = 0.039) were observed in subfertile men (n = 60) compared to fertile men (n = 15). In the whole study group, a positive correlation was also found between the percentage of spermatozoa exhibiting equatorial localization in capacitated samples and normal head forms (R = 0.50; P < 0.0001).     

Anticorps anti-spermatozoïdes: techniques de dépistage et interprétation des résultats

Since the first publication on the detection of sperm-agglutinating antibodies in infertile men, multiple assays have been described. The most useful tests are able to detect antibodies bound to the sperm membrane of motile spermatozoa. The immunobeads test (IBT) is considered to be the most advantageous in terms of its sensitivity, the low incidence of false-positive results, and its ability to localize antibodies of different immunoglobulin classes on the sperm surface. The IBT assay can be used in parallel with the mixed antiglobulin reaction (MAR), to detect sperm-associated antibodies in the ejaculates of infertile men, the most rational way to test for antisperm antibodies (ASA) in males. In view of the high level of agreement between the two assays, MAR, the easier of the two, may be used as a first step in the detection of these antibodies. A positive MAR must be confirmed by IBT, as this assay is more specific for the detection of IgA antibodies. The clinical significance of sperm-associated antibodies is usually established according to the proportion of motile spermatozoa coated with immunobeads, its class and its localization on the sperm surface. However, binding of immunobeads does not provide any information about the antigens against which the antibodies are directed. As the functional effects of sperm-associated antibodies may vary as a function of their antigenic specificities, other assays, using purified fertilization related antigens, are necessary to establish, for each individual, the specific impact of the antibodies on the fertilization process. The indirect IBT assay has recently become the most widely used test to detect the various classes of ASA in serum and cervical mucus of infertile women and in the serum and seminal fluid of infertile men, in combination with the direct assay described above. However, in most laboratories, it is performed with only one dilution of the biological fluid tested, usually a low dilution, so that antibody levels of no significance for fertility could be detected. This may explain a recente debate (Human Reprod, 1999) on the significance of ASA as a cause of infertility. At present, and in the absence of standardized assays able to identify the antigens involved in each individual immune reaction, antibody assays, as detected by the IBT assay, in the serum and/or genital secretions of infertile subjects might provide useful clinical guidance.     

Apoptose et ségrégation méiotique dans les spermatozoïdes d'hommes porteurs de translocations

Men with a chromosomal translocation produce a significant percentage of unbalanced spermatozoa. In order to determine a correlation between chromosomal anomalies and apoptosis in human sperm, we analysed DNA fragmentation and meiotic segregation in sperm from men with a (13;14) Robertsonian translocation. We studied sperm from 12 (13;14) translocation carriers and 9 proven fertile men with a normal karyotype. Meiotic segregation of chromosomes 13 and 14 was analysed using dual-colour fluorescencein situ hybridization with locus-specific probes for chromosomes 13 and 14. Apoptosis in spermatozoa was measured byin situ TUNEL assay. The meiotic segregation study showed a significantly increased frequency of unbalanced spermatozoa for chromosomes 13 and 14 in (13;14) carriers (15.9%) compared to the control population (1.3%) (p=0.00016). The study of apoptosis showed an increase of DNA fragmentation in (13;14) carriers (34.9%) compared to the control population (13.8%) (p=0.0036). This increased apoptosis was observed in spermatozoa presenting an increase of unbalanced chromosomal anomalies concerning chromosomes 13 and 14, but with a predominance of balanced spermatozoa compared to the theoretical risk of meiotic segregation. These results suggest that apoptosis could be involved as a regulatory mechanism to eliminate unbalanced chromosomal spermatozoa in men with a (13;14) Robertsonian translocation.     

Dynamics of a feline retrovirus (FeLV) in host populations with variable spatial structure.

The predictions of epidemic models are remarkably affected by the underlying assumptions concerning host population dynamics and the relation between host density and disease transmission. Furthermore, hypotheses underlying distinct models are rarely tested. Domestic cats (Felis catus) can be used to compare models and test their predictions, because cat populations show variable spatial structure that probably results in variability in the relation between density and disease transmission. Cat populations also exhibit various dynamics. We compare four epidemiological models of Feline Leukaemia Virus (FeLV). We use two different incidence terms, i.e. proportionate mixing and pseudo-mass action. Population dynamics are modelled as logistic or exponential growth. Compared with proportionate mixing, mass action incidence with logistic growth results in a threshold population size under which the virus cannot persist in the population. Exponential growth of host populations results in systems where FeLV persistence at a steady prevalence and depression of host population growth are biologically unlikely to occur. Predictions of our models account for presently available data on FeLV dynamics in various populations of cats. Thus, host population dynamics and spatial structure can be determinant parameters in parasite transmission, host population depression, and disease control.     

Results of epidemiologic studies performed after the disaster in Czernobyl among the adult part of the population in the region of Krakow]

Epidemiologic studies following the Czernobyl accident were performed in region Kraków, including Kraków, Nowy Sacz and Kielce district. 1426 males and 2495 females were selected according to the random sample on the whole population of Kraków and Nowy Sacz, as well as in some selected areas in Swietokrzyski Mountains, and in Kielcecity. The aim of the study was to assess the results of the prophylaxis with Kalium iodine after the radiation and the incidence of the goiter in the population. It was stated, that 19.2% of the population in Kraków district, 16.9% in Nowy Sacz and 20% in Kielce received the prophylactic dosis of K.J. 80% took mainly the Lugol solution, between May, the 1st and 5th, 1986. Among 18 of person showing side effects like gastrointestinal disturbances, 16 were of female sex. Goiter incidence according to WHO classification was 50.7%, 67.3% and 49.9% in Kraków, Nowy Sacz and Kielce respectively. The difference between the incidence of goiter in males and females was 1:3. In women it was rather Ist and IInd degree of goiter, in men OB and Ist. Nodules of thyroid gland in the rural region of Kraków, Nowy Sacz and Kielce were seen in women in 10.8%, 1.7%, add 12.3% consecutively. Hormonal studies i.e T3, T4, TSH serum concentration showed normal results in all groups studied. TSH concentration was the highest in the group OB. The microsomal and antithyroglobulin antibodies level was the same independently on the prophylactic dosis of Lugol solution. The high incidence of thyroid diseases not related to the accident was observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunological structure of the population vis-à-vis the meningococcus]

The immunological and epidemiological study of 3362 serum samples selectively taken from healthy persons in Yaroslavl showed the presence of highly pronounced inverse correlation between morbidity rate indices and level of antibodies, as well as direct correlation between the level of carriership and the level of antibodies, which indicated the specific character of this process. The analysis of serological data showed that all meningococcal antigenic fractions took part in the formation of the immunological structure of the population, but lipopolysaccharides and polysaccharides played the most active role in this process. The level of all immunoglobulin classes (especially IgA) in the sera of the persons selected for this study was also shown to depend on their age.     

Prevalence of antibodies to Francisella tularensis in forest workers from different regions of Poland

In the present study we evaluate the prevalence of antibodies to F. tularensis in 480 serum samples obtained from healthy forest workers from different regions of Poland. The investigations were performed using the tube agglutination test and ELISA. The cut-off limit of serum antibodies was set at mean antibody titre determined in the sera of 115 blood donors exceeded by three standard deviations. In none serum samples we detected antibodies to F. tularensis by tube agglutination test. Of the 480 tested sera IgA antibodies were detected by ELISA in 4.6%, antibodies IgG in 3.8% and antibodies IgM in 2.70% serum samples. The results of our study showed that antibodies to F. tularensis were slightly, but not statistically significant, more often diagnosed in healthy forest workers than healthy blood donors.