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Association between levels of persistent organic pollutants in adipose tissue and cryptorchidism in early childhood: a case-control study.

https://arctichealth.org/en/permalink/ahliterature272722
Source
Environ Health. 2015;14:78
Publication Type
Article
Date
2015
Author
Jaakko J Koskenniemi
Helena E Virtanen
Hannu Kiviranta
Ida N Damgaard
Jaakko Matomäki
Jørgen M Thorup
Timo Hurme
Niels E Skakkebaek
Katharina M Main
Jorma Toppari
Source
Environ Health. 2015;14:78
Date
2015
Language
English
Publication Type
Article
Keywords
Adipose Tissue - chemistry
Benzofurans - toxicity
Case-Control Studies
Child
Child, Preschool
Cross-Sectional Studies
Cryptorchidism - chemically induced - epidemiology
Denmark - epidemiology
Dioxins - toxicity
Environmental Pollutants - toxicity
Female
Finland - epidemiology
Halogenated Diphenyl Ethers - toxicity
Humans
Incidence
Infant
Infant, Newborn
Male
Polychlorinated Biphenyls - toxicity
Pregnancy
Prenatal Exposure Delayed Effects - chemically induced - epidemiology
Abstract
Congenital cryptorchidism, i.e. failure of the testicular descent to the bottom of the scrotum, is a common birth defect. The evidence from epidemiological, wildlife, and animal studies suggests that exposure to mixtures of endocrine disrupting chemicals during fetal development may play a role in its pathogenesis. We aimed to assess the association between cryptorchidism and prenatal exposure to polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs), and polybrominated diphenyl ethers (PBDEs).
We conducted a case-control study consisting of 44 cryptorchid cases, and 38 controls operated for inguinal hernia, umbilical hernia, or hydrocele at the Turku University Hospital or Rigshospitalet, Copenhagen in 2002-2006. During the operation a subcutaneous adipose tissue biopsy was taken. Samples were analysed for 37 PCBs, 17 PCDD/Fs and 14 PBDEs by gas chromatography-high-resolution mass spectrometry. Chemical concentrations were adjusted for postnatal variation introduced by differences in duration of breastfeeding, age at the operation, and country of origin with a multiple linear regression. Association between adjusted and unadjusted chemical concentrations and the risk of cryptorchidism were analysed with logistic regression to get an estimate for odds ratio (OR) of cryptorchidism per multiplication of chemical concentrations with ca. 2.71 (Napier's constant).
Total-TEq i.e. the WHO-recommended 2,3,7,8-TCDD equivalent quantity of 17 dioxins and 12 dioxin-like PCBs and sum of PCDD/Fs were positively associated with cryptorchidism [OR 3.21 (95% CI 1.29-9.09), OR 3.69 (95% CI 1.45-10.9), respectively], when adjusting for country of origin, the duration the child was breastfed, and age at operation. The association between the sum of PCBs and cryptorchidism was close to significant [OR 1.92 (95% CI 0.98-4.01)], whereas the association between the sum of PBDEs and cryptorchidism was not [OR 0.86 (95% CI 0.47-1.54)]. There were no associations between unadjusted chemical concentrations and the risk of cryptorchidism.
Prenatal exposure to PCDD/Fs and PCDD/F-like PCBs may be associated with increased risk for cryptorchidism. Our finding does not exclude the possibility of an association between the exposure to PBDEs and cryptorchidism.
Notes
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PubMed ID
26403566 View in PubMed
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Association of placenta organotin concentrations with congenital cryptorchidism and reproductive hormone levels in 280 newborn boys from Denmark and Finland.

https://arctichealth.org/en/permalink/ahliterature115317
Source
Hum Reprod. 2013 Jun;28(6):1647-60
Publication Type
Article
Date
Jun-2013
Author
Panu Rantakokko
Katharina M Main
Christine Wohlfart-Veje
Hannu Kiviranta
Riikka Airaksinen
Terttu Vartiainen
Niels E Skakkebæk
Jorma Toppari
Helena E Virtanen
Author Affiliation
National Institute for Health and Welfare, Department of Environmental Health, Chemical Exposure Unit, Neulaniementie 4, FI-70210 Kuopio, Finland. panu.rantakokko@thl.fi
Source
Hum Reprod. 2013 Jun;28(6):1647-60
Date
Jun-2013
Language
English
Publication Type
Article
Keywords
Adult
Cohort Studies
Cryptorchidism - etiology
Denmark
Endocrine Disruptors - metabolism
Female
Finland
Follicle Stimulating Hormone - blood
Gas Chromatography-Mass Spectrometry
Gonadotropins, Pituitary - blood
Humans
Infant, Newborn
Logistic Models
Luteinizing Hormone - blood
Male
Organotin Compounds - metabolism
Placenta - metabolism
Pregnancy
Prenatal Exposure Delayed Effects
Risk factors
Sex Hormone-Binding Globulin - metabolism
Trialkyltin Compounds - metabolism
Abstract
Is the placental burden of organotin compounds (OTCs) associated with congenital cryptorchidism in infant offspring from Finland and Denmark?
Increasing concentrations of OTCs had a negative association with cryptorchidism in Finland, whereas a positive association was found in Denmark.
The rapid increase in the prevalence of cryptorchidism suggests that environmental factors, such as endocrine disruptors, may be involved. OTCs are endocrine disruptors at very low concentrations due to activation of the retinoid X receptor (RXR).
Between the years 1997 and 2001, placentas from mothers of cryptorchid boys and from healthy controls were collected from Denmark (39 cases, 129 controls) and Finland (56 cases, 56 controls). In Denmark 33 and 6 boys, and in Finland 22 and 34 boys had mild or severe cryptorchidism, respectively. The association between concentrations of four OTCs [monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT) and triphenyltin (TPhT)] and case-control status was estimated.
In both countries, placenta samples were selected from larger cohorts. In Finland placenta samples were collected from boys with cryptorchidism at birth and matched controls (nested case-control design). Matching criteria were parity, maternal smoking (yes/no), diabetes (yes/no), gestational age (±7 days) and date of birth (±14 days). Numbers of controls per case was 1. In Denmark, all available placentas from cryptorchid boys were chosen and control placentas were selected randomly from the total Danish cohort (case-cohort design). The average number of controls per case was 3.3. OTCs in placenta samples were analysed with liquid extraction, ethylation and gas chromatography-mass spectrometry determination and coded by country-specific tertiles.
Generally, the concentrations of OTCs were very low. For most analytes, a large proportion of samples (29-96% depending on the country and case-control status) had OTC concentrations below the limit of quantification (LOQ). As an exception, the concentration of TBT was >LOQ in 99% of Finnish placentas. The mean concentrations of DBT and TBT were 1.5 and 7 times higher in Finland than in Denmark, respectively. For DBT in Danish placentas, the odds ratio (OR) for cryptorchidism in the second tertile (0.10-0.14 ng/g) when compared with the first tertile (
PubMed ID
23520400 View in PubMed
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Association of placenta organotin concentrations with growth and ponderal index in 110 newborn boys from Finland during the first 18 months of life: a cohort study.

https://arctichealth.org/en/permalink/ahliterature257106
Source
Environ Health. 2014;13(1):45
Publication Type
Article
Date
2014
Author
Panu Rantakokko
Katharina M Main
Christine Wohlfart-Veje
Hannu Kiviranta
Riikka Airaksinen
Terttu Vartiainen
Niels E Skakkebæk
Jorma Toppari
Helena E Virtanen
Author Affiliation
National Institute for Health and Welfare, Department of Environmental Health, Toxicology and Chemical Exposure Unit, Neulaniementie 4, FI-70210 Kuopio, Finland. panu.rantakokko@thl.fi.
Source
Environ Health. 2014;13(1):45
Date
2014
Language
English
Publication Type
Article
Keywords
Adult
Child Development
Cohort Studies
Cryptorchidism - epidemiology
Environmental Pollutants - analysis
Female
Finland - epidemiology
Humans
Infant, Newborn
Male
Organotin Compounds - analysis
Placenta - chemistry
Pregnancy
Weight Gain
Young Adult
Abstract
Humans are exposed to tributyltin (TBT), previously used as an antifouling paint in ships, mainly through fish consumption. As TBT is a known obesogen, we studied the association of placenta TBT and other organotin compounds (OTCs) with ponderal index (PI) and growth during the first 18 months of life in boys.
In a prospective Finnish study, 110 placenta samples were collected from mothers of boys born in 1997-1999 with (n?=?55) and without (n?=?55) cryptorchidism. To account for the original study design, linear regression, weighted for sampling fractions of boys with (121/55) and without (5677/55) cryptorchidism from the total cohort, was used to study the association between placenta OTCs and children's weight, length, growth rates and PI up to 18 months of age.
Placenta TBT concentrations were above the limit of quantification (LOQ) in 99% of the samples. However, monobutyltin (MBT), dibutyltin (DBT) and triphenyltin (TPhT) concentrations were below LOQ in 90%, 35% and 57% of samples, respectively. Placenta TBT was positively associated (p?=?0.024) with weight gain during the first three months of life, but no other significant associations were observed for weight or length gain. Also, no significant associations between placenta OTC concentrations and child length, weight or PI at any time point were found.
We observed a trend towards higher weight gain from birth to 3 months of age with increasing placenta TBT concentration. These results should be interpreted with caution because obesogenic effects in animal experiments were seen after in-utero TBT exposures to doses that were orders of magnitude higher. Also the number of study subjects included in this study was limited.
Notes
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PubMed ID
24899383 View in PubMed
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Circannual rhythm in the incidence of cryptorchidism in Finland.

https://arctichealth.org/en/permalink/ahliterature176412
Source
Int J Androl. 2005 Feb;28(1):53-7
Publication Type
Article
Date
Feb-2005
Author
Marko Kaleva
Helena E Virtanen
Anne-Maarit Haavisto
Katharina M Main
Mikko Reunanen
Niels E Skakkebaek
Jorma Toppari
Author Affiliation
Department of Physiology, University of Turku, Kiinamyllynkatu 10, FIN-20520 Turku, Finland. marko.kaleva@utu.fi
Source
Int J Androl. 2005 Feb;28(1):53-7
Date
Feb-2005
Language
English
Publication Type
Article
Keywords
Circadian Rhythm
Cryptorchidism - epidemiology
Finland - epidemiology
Humans
Incidence
Male
Abstract
Conflicting data on circannual variation in birth rates of urogenital malformations have been reported previously. To assess risk factors of cryptorchidism we studied the seasonal variation of cryptorchidism in Finland. We performed a prospective cryptorchidism study in Turku, Finland, from 1997 to 2001 to evaluate the incidence of cryptorchidism. Clinical examinations were performed at birth and at 3 months. Of 9511 liveborn boys (1471 preterm boys) 216 (53 preterm boys) were cryptorchid at birth and 106 (19 preterm boys) at 3 months. The incidence of cryptorchidism was significantly higher in spring (February-April) (3.0%) than in summer (May-July) (1.7%) (OR 1.79; 95% CI: 1.23-2.63). This seasonal difference was observed both among preterm and term boys. We conclude that a circannual fluctuation in the incidence of cryptorchidism exists, which indicates an influence by environmental factors. The underlying reason for cyclicity affects similarly both preterm and term boys.
PubMed ID
15679622 View in PubMed
Less detail

Genital anomalies in boys and the environment.

https://arctichealth.org/en/permalink/ahliterature142933
Source
Best Pract Res Clin Endocrinol Metab. 2010 Apr;24(2):279-89
Publication Type
Article
Date
Apr-2010
Author
Katharina M Main
Niels E Skakkebaek
Helena E Virtanen
Jorma Toppari
Author Affiliation
University Department of Growth and Reproduction GR, Section 5064, Rigshospitalet, Blegdamsvej 9, DK-2100 Copenhagen, Denmark. katharina.main@rh.regionh.dk
Source
Best Pract Res Clin Endocrinol Metab. 2010 Apr;24(2):279-89
Date
Apr-2010
Language
English
Publication Type
Article
Keywords
Cryptorchidism - complications - epidemiology
Denmark - epidemiology
Endocrine Disruptors - toxicity
Environment
Environmental Pollutants - toxicity
Female
Finland - epidemiology
Genitalia, Male - abnormalities - drug effects
Gonadal Dysgenesis
Halogenated Diphenyl Ethers - toxicity
Humans
Hypospadias - epidemiology
Infant, Newborn
Male
Milk, human - chemistry
Phthalic Acids - metabolism
Pregnancy
Testicular Neoplasms - epidemiology
Testis - abnormalities
Abstract
The prevalence of male reproductive disorders, such as testicular cancer and impaired semen quality, is increasing in many, albeit not all, countries. These disorders are aetiologically linked with congenital cryptorchidism and hypospadias by common factors leading to perinatal disruption of normal testis differentiation, the testicular dysgenesis syndrome (TDS). There is recent evidence that also the prevalence of genital malformations is increasing and the rapid pace of increase suggests that lifestyle factors and exposure to environmental chemicals with endocrine disrupting properties may play a role. Recent prospective studies have established links between perinatal exposure to persistent halogenated compounds and cryptorchidism, as well as between phthalates and anti-androgenic effects in newborns. Maternal alcohol consumption, mild gestational diabetes and nicotine substitutes were also identified as potential risk factors for cryptorchidism. It may be the cocktail effect of many simultaneous exposures that result in adverse effects, especially during foetal life and infancy.
PubMed ID
20541152 View in PubMed
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Human breast milk contamination with phthalates and alterations of endogenous reproductive hormones in infants three months of age.

https://arctichealth.org/en/permalink/ahliterature76061
Source
Environ Health Perspect. 2006 Feb;114(2):270-6
Publication Type
Article
Date
Feb-2006
Author
Katharina M Main
Gerda K Mortensen
Marko M Kaleva
Kirsten A Boisen
Ida N Damgaard
Marla Chellakooty
Ida M Schmidt
Anne-Maarit Suomi
Helena E Virtanen
Dajørgen Vid H Petersen
Anna-Maria Andersson
Jorma Toppari
Niels E Skakkebaek
Author Affiliation
University Department of Growth and Reproduction, Rigshospitalet, Copenhagen, Denmark. katharina.main@rh.hosp.dk
Source
Environ Health Perspect. 2006 Feb;114(2):270-6
Date
Feb-2006
Language
English
Publication Type
Article
Keywords
Adult
Cohort Studies
Cryptorchidism - chemically induced
Denmark
Female
Finland
Gonadal Steroid Hormones - blood
Humans
Infant, Newborn
Longitudinal Studies
Male
Milk, human - chemistry
Phthalic Acids - analysis - poisoning
Pregnancy
Prenatal Exposure Delayed Effects
Research Support, Non-U.S. Gov't
Abstract
Phthalates adversely affect the male reproductive system in animals. We investigated whether phthalate monoester contamination of human breast milk had any influence on the postnatal surge of reproductive hormones in newborn boys as a sign of testicular dysgenesis. DESIGN: We obtained biologic samples from a prospective Danish-Finnish cohort study on cryptorchidism from 1997 to 2001. We analyzed individual breast milk samples collected as additive aliquots 1-3 months postnatally (n = 130; 62 cryptorchid/68 healthy boys) for phthalate monoesters [mono-methyl phthalate (mMP), mono-ethyl phthalate (mEP), mono-n-butyl phthalate (mBP), mono-benzyl phthalate (mBzP), mono-2-ethylhexyl phthalate (mEHP), mono-isononyl phthalate (miNP)]. We analyzed serum samples (obtained in 74% of all boys) for gonadotropins, sex-hormone binding globulin (SHBG), testosterone, and inhibin B. RESULTS: All phthalate monoesters were found in breast milk with large variations [medians (minimum-maximum)]: mMP 0.10 (
PubMed ID
16451866 View in PubMed
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Larger Testes and Higher Inhibin B Levels in Finnish than in Danish Newborn Boys.

https://arctichealth.org/en/permalink/ahliterature16377
Source
J Clin Endocrinol Metab. 2006 Apr 4;
Publication Type
Article
Date
Apr-4-2006
Author
Katharina M Main
Jorma Toppari
Anne-Maarit Suomi
Marko Kaleva
Marla Chellakooty
Ida M Schmidt
Helena E Virtanen
Kirsten A Boisen
Claudia M Kai
Ida N Damgaard
Niels E Skakkebæk
Author Affiliation
University Department of Growth and Reproduction, Rigshospitalet, Copenhagen, Denmark; Departments of Physiology and Paediatrics, University of Turku, Finland.
Source
J Clin Endocrinol Metab. 2006 Apr 4;
Date
Apr-4-2006
Language
English
Publication Type
Article
Abstract
Context: Recent studies showed that male reproductive health problems like cryptorchidism, hypospadias, testicular cancer and low sperm quality are more prevalent in Denmark than in Finland. Objectives: We hypothesized that if fetal testicular dysgenesis contributed to these observations, differences in gonadal development and the hypothalamus-pituitary-testis axis would already be detectable perinatally. Thus, we investigated healthy newborn boys in both countries. Design: Prospective, longitudinal population-based study. Setting: Two primary obstetric centers at the University Hospitals of Copenhagen, Denmark, and Turku, Finland Participants: 633 Danish / 1044 Finnish boys, born at term with appropriate weight for gestational age Interventions: ultrasound determination of testis size at 0-3-18 months and blood sampling (n = 727) at 3 months Main outcome measures: testicular volume, reproductive hormones Results: Testis volume was significantly higher at all ages in Finnish than in Danish boys (medians: 98 vs. 95 mm(3), 185 vs. 119 mm(3) and 188 vs. 136 mm(3), respectively, P
PubMed ID
16595596 View in PubMed
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No association between exposure to perfluorinated compounds and congenital cryptorchidism: a nested case-control study among 215 boys from Denmark and Finland.

https://arctichealth.org/en/permalink/ahliterature259742
Source
Reproduction. 2014;147(4):411-7
Publication Type
Article
Date
2014
Author
Dorte Vesterholm Jensen
Jeppe Christensen
Helena E Virtanen
Niels E Skakkebæk
Katharina M Main
Jorma Toppari
Christine W Veje
Anna-Maria Andersson
Flemming Nielsen
Philippe Grandjean
Tina Kold Jensen
Source
Reproduction. 2014;147(4):411-7
Date
2014
Language
English
Publication Type
Article
Keywords
Adult
Alkanesulfonic Acids - blood
Caprylates - blood
Case-Control Studies
Cryptorchidism - blood - chemically induced - epidemiology
Denmark - epidemiology
Endocrine Disruptors - blood
Female
Fetal Blood - chemistry
Finland - epidemiology
Fluorocarbons - blood
Humans
Infant, Newborn
Male
Pregnancy
Prenatal Exposure Delayed Effects - blood - epidemiology
Abstract
Geographical differences in the occurrence of diseases in male reproductive organs, including malformation in reproductive tract, between Denmark and Finland have been reported. The reason for these differences is unknown, but differences in exposure to chemicals with endocrine-disrupting abilities have been suggested. Among these chemicals are perfluoro-alkylated substances (PFASs), a group of water- and grease-repellent chemicals used in outdoor clothes, cookware, food packaging, and textiles. In this study, we, therefore, investigated differences in PFAS exposure levels between Denmark and Finland and the association between cord blood PFAS levels and congenital cryptorchidism. Boys from a joint ongoing prospective birth cohort study were included. We analyzed PFAS levels in cord blood serum samples collected from 29 Danish boys with congenital cryptorchidism, 30 healthy Danish matched controls recruited from 1997 to 2001, 30 Finnish cases, and 78 Finnish healthy matched controls recruited from 1997 to 1999. Additionally, 48 Finnish cases recruited from 2000 to 2002 were included. Perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) were detected in all the 215 Danish and Finnish cord blood samples with significantly higher levels being observed in the Danish samples (medians: PFOA, 2.6?ng/ml and PFOS, 9.1?ng/ml) than in the Finnish samples (medians: PFOA, 2.1?ng/ml and PFOS, 5.2?ng/ml). We found no associations between cord blood PFOA and PFOS levels and congenital cryptorchidism after adjustment for confounders. Our data indicate that women in Denmark and Finland are generally exposed to PFOA and PFOS but there are differences in exposure levels between countries. We found no statistically significant association between cord blood PFOA and PFOS levels and congenital cryptorchidism; however, our study was small and larger studies are warranted.
PubMed ID
24218628 View in PubMed
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Postnatal penile length and growth rate correlate to serum testosterone levels: a longitudinal study of 1962 normal boys.

https://arctichealth.org/en/permalink/ahliterature29290
Source
Eur J Endocrinol. 2006 Jan;154(1):125-9
Publication Type
Article
Date
Jan-2006
Author
Malene Boas
Kirsten A Boisen
Helena E Virtanen
Marko Kaleva
Anne-Maarit Suomi
Ida M Schmidt
Ida N Damgaard
Claudia M Kai
Marla Chellakooty
Niels E Skakkebaek
Jorma Toppari
Katharina M Main
Author Affiliation
University Department of Growth and Reproduction Section 5064, Rigshospitalet, Blegdamsvej 9, DK-2100 Copenhagen, Denmark.
Source
Eur J Endocrinol. 2006 Jan;154(1):125-9
Date
Jan-2006
Language
English
Publication Type
Article
Keywords
Aging
Child, Preschool
Denmark
Finland
Humans
Infant
Infant, Newborn
Longitudinal Studies
Male
Penis - anatomy & histology - growth & development
Reference Values
Research Support, Non-U.S. Gov't
Testosterone - blood
Abstract
OBJECTIVE: Infant boys show a brief activation of their hypothalamic-pituitary-gonadal axis shortly after birth, the physiological significance of which is poorly understood. The objective of the study was to investigate the correlation between endogenous testosterone levels and penile size and growth. DESIGN: Prospective, longitudinal population-based study taking place at two large primary obstetric centres at the University Hospitals of Copenhagen, Denmark, and Turku, Finland. METHODS: Infant boys, 728 Danish and 1234 Finnish, underwent clinical examinations at 0, 3, 18 and 36 months in Denmark and at 0, 3 and 18 months in Finland with blood samples taken at 3 months (n = 630). Penile length and growth were registered and reproductive hormones (testosterone, sex hormone binding globulin, oestradiol) were analysed. RESULTS: Penile length increased from birth (3.49+/-0.4 cm) to 3 years of age (4.53+/-0.51 cm) with the highest growth velocity from birth to 3 months (1.0 mm/month). Penile length and growth were significantly, positively correlated to serum testosterone (r = 0.31 and 0.076, P = 0.006 and 0.001 respectively) and to free testosterone index (r = 0.385 and 0.094, P = 0.0001 and 0.0001 respectively). CONCLUSIONS: We found that endogenous testosterone was significantly associated with penile size and growth rate in infant boys. Thus, the postnatal surge in reproductive hormones appears to be important for genital growth. Our data may serve as an updated reference for normal penile length in Caucasian boys up to 3 years of age.
PubMed ID
16382001 View in PubMed
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Semen quality improves marginally during young adulthood: a longitudinal follow-up study.

https://arctichealth.org/en/permalink/ahliterature278136
Source
Hum Reprod. 2016 Mar;31(3):502-10
Publication Type
Article
Date
Mar-2016
Author
Antti Perheentupa
Sergey Sadov
Riitta Rönkä
Helena E Virtanen
Wiwat Rodprasert
Matti Vierula
Niels Jørgensen
Niels E Skakkebæk
Jorma Toppari
Source
Hum Reprod. 2016 Mar;31(3):502-10
Date
Mar-2016
Language
English
Publication Type
Article
Keywords
Adult
Age Factors
Finland
Humans
Longitudinal Studies
Male
Multivariate Analysis
Reproductive health
Semen Analysis
Sperm Count
Sperm Motility
Spermatogenesis
Abstract
Does semen quality improve during early adulthood?
Semen variables change little during the third decade of life, however some improvement in sperm morphology and motility may occur.
A suspicion of deteriorating semen quality has been raised in several studies. The longitudinal development of semen quality in early adulthood is insufficiently understood.
A longitudinal follow-up of two cohorts of volunteer young adult Finnish men representing the general population was carried out. Cohorts A (discovery cohort, born 1979-1981, n = 336) and B (validation cohort, born 1983, n = 197) were followed up from the age of 19 years onward for 10 years.
Inclusion criteria included that both the men and their mothers were born in Finland. Semen analysis was performed in cohorts A and B at 2-4 year intervals over a period of 10 years. Semen volume, sperm concentration, total sperm count, motility, total motile count and morphology were the variables assessed in the analysis. A physical examination was carried out at each visit to detect any significant andrological abnormalities. The overall participation rate was 13.4%.
During the follow-up, the percentage of sperm with normal morphology and the percentage of motile sperm increased significantly both in the discovery (A) (P
Notes
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PubMed ID
26740579 View in PubMed
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