Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 5 uppsatser.

Cryptorchidism is a male congenital disorder with an unspecified, multifactorial etiology. This review evaluated the strength of select factors in the development of cryptorchidism to better understand its etiology. The strength of relationship between factors and their respective functions during testicular descent was evaluated. Factors evaluated in the causal pathway include the signaling mechanisms Desert Hedgehog (DHH), Insulin-like Hormone 3 (INSL3) and Platelet-Derived Growth Factor (PDGF), as well as sex hormone regulation (androgen: estrogen ratio, aromatase expression). Articles supporting a factor in testicular descent were evaluated and scored. These scores were summed to create the “Step Score” for each step in the causal pathway. An arrow system was developed which ranked the strength of each pathway step as either “weak”, “moderate” or “strong”. Thus, step scores and the strength of factors in the pathological pathway were determined: DHH (15-moderate), PDGF (10-weak), INSL3 (24-strong) and Androgen: Estrogen ratio, Aromatase (23-strong). The pathological pathway produced by this review represents a literature based perspective of the research regarding cryptorchidism etiology. Literature indicates that prenatal exposure to endocrine disrupting chemicals in animals and humans may lead to abnormal genital development. Recently, prenatal maternal cigarette smoke was demonstrated to be a risk factor for cryptorchidism. This controversial finding was explored in the context of endocrine disrupting chemicals. However, literature has provided very little evidence in support of this hypothesis and more research is needed to better evaluate prenatal maternal smoking as a risk factor for undescended testis.

O objetivo deste trabalho é estudar a incidência, a estrutura dos apêndices testiculares (AT) em pacientes com criptorquia, comparando a sua incidência com as anomalias epididimárias e a patência do processo vaginal. Estudamos 55 pacientes (72 testículos) portadores de criptorquia e 8 testículos como controle (6 hidroceles e 2 torções). Analisamos as relações entre o testículo e o epidídimo, a patência do processo vaginal e a incidência e histologia dos ATs. Fibras musculares lisas (SMC), tecido conectivo (CT) e fibras do sistema elástico (ESF) foram estudados por métodos imuno-histoquímicos. Dos 72 testículos com criptorquia 20 (27.77%) apresentavam anomalias epididimárias, 41(56.9%) tinham processo vaginal patente e 44 (61.1%) tinham apêndices testiculares. Dos 44 testículos portadores de criptorquia com apêndices, 30 (68.18%) apresentavam o processo vaginal patente e 11 (25%) apresentavam anomalias epididimárias. O epitélio não apresentou alteração aparente nos apêndices de pacientes com criptorquia e no grupo controle. Análise estereológica documentou a prevalência de ESF (média de 1.48%); prevalência de vasos (média de 10,11%) e uma diminuição (p=0.14) da SMC nos AT de pacientes com criptorquia (média = 4.93%). O colágeno III prevaleceu nos AT de pacientes com criptorquia. Não houve alteração na incidência de anomalias anatômicas associadas aos testículos portadores de AT. Os apêndices testiculares apresentaram uma alteração estrutural significativa nos pacientes com criptorquia, o que indica que os AT apresentam uma remodelação estrutural significativa nos pacientes com criptorquia. / This work reports the incidence and structure of testicular appendices (TAs) in patients with cryptorchidism, comparing their incidence with epididymal anomalies and patency of the vaginal process. We studied 55 patients (72 testes) with cryptorchidism and 8 testes as controls (6 with hydroceles and 2 with torsion). We analyzed the relations among the testis, epididymis and patency of the vaginal process and prevalence and histology of the TAs. Smooth muscle cells (SMCs), connective tissue (CT) and elastic system fibers (ESFs) were studied by histochemical and immunolabeling methods. Of the 72 testes with cryptorchidism, 20 (27.77%) presented epididymal anomalies, 41 (56.9%) had patent vaginal processes and 44 (61.1%) had TAs. Of the 44 testes with cryptorchidism and appendices, 30 (68.18%) presented patent vaginal processes and 11 (25%) presented epididymal anomalies. There was no apparent alteration of the epithelium in the appendices of patients with cryptorchidism and in the control group. Stereological analysis documented the prevalence of ESFs (mean of 1.48%), prevalence of veins (mean of 10.11%) and decrease (p=0.14) of SMCs in the TAs of patients with cryptorchidism (mean = 4.93%). Collagen III prevailed in the TAs of patients with cryptorchidism. There was no alteration in the incidence of anatomical anomalies associated with the testes with TAs. The testicular appendices presented significant structural alteration in the patients with cryptorchidism, indicating that TAs present a significant structural remodeling in patients with cryptorchidism.

O objetivo deste trabalho é estudar a incidência, a estrutura dos apêndices testiculares (AT) em pacientes com criptorquia, comparando a sua incidência com as anomalias epididimárias e a patência do processo vaginal. Estudamos 55 pacientes (72 testículos) portadores de criptorquia e 8 testículos como controle (6 hidroceles e 2 torções). Analisamos as relações entre o testículo e o epidídimo, a patência do processo vaginal e a incidência e histologia dos ATs. Fibras musculares lisas (SMC), tecido conectivo (CT) e fibras do sistema elástico (ESF) foram estudados por métodos imuno-histoquímicos. Dos 72 testículos com criptorquia 20 (27.77%) apresentavam anomalias epididimárias, 41(56.9%) tinham processo vaginal patente e 44 (61.1%) tinham apêndices testiculares. Dos 44 testículos portadores de criptorquia com apêndices, 30 (68.18%) apresentavam o processo vaginal patente e 11 (25%) apresentavam anomalias epididimárias. O epitélio não apresentou alteração aparente nos apêndices de pacientes com criptorquia e no grupo controle. Análise estereológica documentou a prevalência de ESF (média de 1.48%); prevalência de vasos (média de 10,11%) e uma diminuição (p=0.14) da SMC nos AT de pacientes com criptorquia (média = 4.93%). O colágeno III prevaleceu nos AT de pacientes com criptorquia. Não houve alteração na incidência de anomalias anatômicas associadas aos testículos portadores de AT. Os apêndices testiculares apresentaram uma alteração estrutural significativa nos pacientes com criptorquia, o que indica que os AT apresentam uma remodelação estrutural significativa nos pacientes com criptorquia. / This work reports the incidence and structure of testicular appendices (TAs) in patients with cryptorchidism, comparing their incidence with epididymal anomalies and patency of the vaginal process. We studied 55 patients (72 testes) with cryptorchidism and 8 testes as controls (6 with hydroceles and 2 with torsion). We analyzed the relations among the testis, epididymis and patency of the vaginal process and prevalence and histology of the TAs. Smooth muscle cells (SMCs), connective tissue (CT) and elastic system fibers (ESFs) were studied by histochemical and immunolabeling methods. Of the 72 testes with cryptorchidism, 20 (27.77%) presented epididymal anomalies, 41 (56.9%) had patent vaginal processes and 44 (61.1%) had TAs. Of the 44 testes with cryptorchidism and appendices, 30 (68.18%) presented patent vaginal processes and 11 (25%) presented epididymal anomalies. There was no apparent alteration of the epithelium in the appendices of patients with cryptorchidism and in the control group. Stereological analysis documented the prevalence of ESFs (mean of 1.48%), prevalence of veins (mean of 10.11%) and decrease (p=0.14) of SMCs in the TAs of patients with cryptorchidism (mean = 4.93%). Collagen III prevailed in the TAs of patients with cryptorchidism. There was no alteration in the incidence of anatomical anomalies associated with the testes with TAs. The testicular appendices presented significant structural alteration in the patients with cryptorchidism, indicating that TAs present a significant structural remodeling in patients with cryptorchidism.

Leydig cells produce testosterone in response to luteinizing hormone (LH) via the cyclic adenosine monophosphate (cAMP)/protein kinase A pathway. Additionally, these cells are responsible for producing insulin-like peptide 3 (INSL3), a peptide hormone that is essential for testicular descent. The insl3 promoter in Leydig cells can be activated by cAMP through the transcription factor Nur77, which has also been shown to regulate the promoters of the steroidogenic enzymes, cyp17 and 3b-hsd. While the mechanism of LH action on testosterone production is well characterized, the effect of LH on insl3 abundance has yet to be shown directly. The MA-10 Leydig cells treated with hCG exhibited a transient and robust increase in nur77 mRNA, while insl3 mRNA abundance remained unchanged. Further, cAMP failed to affect insl3 mRNA, though nur77 mRNA abundance was significantly increased. Inhibition of LH-receptor-linked signal transduction pathways in the presence of hCG implicated multiple signaling networks in the regulation of both insl3 and nur77. Treatment with hCG or cAMP did not affect the abundance of 3b-hsd mRNA. Interestingly, though the MA-10 cell line has been reported to lack CYP17 activity and mRNA and so produce progesterone instead of testosterone, cyp17 mRNA was present and inducible by hCG and cAMP. The addition of hCG, testosterone, nor the combination of hCG and testosterone affected insl3 mRNA abundance. Though hCG consistently increased nur77 mRNA abundance, the addition of testosterone did not enhance the effects of hCG. Collectively, these results indicate that insl3 is regulated by factors other than LH/CG and cAMP in the MA-10 cell line.

One performed a more modern approach on such heat stress to find changes out to compact morphometry in spermatozoon head by computational analysis. One aimed in this work to assess how to compact chromatin and sperm morphometry after have risen testicular temperature, by using experimental cryptorchidism in rabbits (Oryctolagus cuniculus), as well as used 30 rabbits: wild rabbit, white, adult, located in the rabbit keeping of University of Uberaba, Uberaba (city), MG (State of Minas Gerais), Brazil. One classified the rabbits into two groups of 15. Both groups were anesthetised, but only the experimental cryptorchidism one underwent a surgical operation. One gathered semen samples within a total of 69 days. One both dyed semen smear of all rabbits by using a toluidine blue dye and afterwards analysed it with a digital microcomputer to assess compacting chromatin and sperm morphometry. Chromatin varied regarding compact intensity (Diff%), homogeneity by coefficient of variation (CV) and aberrant chromatin (AbChr). By analysing morphometrically one assessed: area, perimeter (Perim.), width (w.), length (l.), ratio width:length (w./l.), ellipsity (E.), form factor (FF), lateral symmetry (LatSym), antero-posterior symmetry (A-PSym), and Fourier descriptors with amplitude from 0 to 2. Testicular temperature rise swayed the concentration, motility, and vigour adversely. Chromatin structure becomes aberrant when affected by testicular heat stress. And spermatozoon head tends to lessen. / Uma abordagem mais moderna sobre a elevação da temperatura foi realizada, visando detectar alterações na compactação e morfometria da cabeça do espermatozoide pela análise computacional. O objetivo do trabalho foi avaliar a compactação da cromatina e a morfometria espermática após a elevação da temperatura testicular, por meio de criptorquidismo experimental, em coelhos. Foram utilizados 30 coelhos da raça Nova Zelândia, brancos, adultos, situados no coelhário da Universidade de Uberaba, Uberaba, MG, Brasil. Os coelhos foram separados em dois grupos de 15. Ambos os grupos foram anestesiados, mas somente um deles sofreu intervenção cirúrgica, o criptorquidismo experimental. Foram coletadas amostras de sêmen durante 69 dias. Esfregaços de sêmen de todos os coelhos foram corados com azul de toluidina e, posteriormente, analisados digitalmente por meio de microcomputador para avaliar a compactação de cromatina e morfometria. A cromatina foi avaliada quanto à intensidade de compactação (Dif%), homogeneidade pelo coeficiente de variação (CV) e cromatina anômala (Cr. An.). Na análise morfométrica foram avaliados: área, perímetro (Per.), largura (Larg.), comprimento (Comp.), razão largura:comprimento (L/C), elipsidade (E), fator forma (FF), simetria lateral (Sim. Lat.), simetria ântero posterior (Sim. A-P) e descritores Fourier com amplitude de 0 a 2. A elevação da temperatura testicular influenciou negativamente a concentração, motilidade, vigor, torna-se anômala a estrutura da cromatina e a cabeça do espermatozoide tende a diminuir. / Mestre em Ciências Veterinárias

Univerzita Karlova v Praze Přírodovědecká fakulta Studijní program: Vývojová biologie (P1520) Studijní obor: Vývojová biologie (1501V000) Oleksandr Chernyavskiy Zdokonalené metody pro snímání obrazových dat a analýzu tkání a buněk pomocí konfokální a multifotonové mikroskopie Improved Methods of Image Acquisition and Analysis of Tissues and Cells by Confocal and Multi-Photon Microscopy Abstrakt disertační práce Školitel: RNDr. Lucie Kubínová CSc Praha, 2015 Abstract The aim of this study was to develop methods and approaches for image acquisition with subsequent image analysis of data, obtained by confocal and two- photon excitation microscopy as well as their combination, enabling new possibilities of visualization and assessment of information on biological tissues and cell structures in 3D and their measurement. We focused on methods that exploited advantages of confocal and multi-photon excitation microscopy. Our further aim was to demonstrate the applicability of non-invasive approach for in vivo applications, usefulness and the relevance of these methods in several special biological applications with emphasis on improved image acquisition, analysis and evaluation of real biological specimens. The present work was not oriented on just one specific biological problem, but rather to methodological...

O hormônio anti-Mülleriano (AMH) é um marcador de reserva ovariana e função testicular. A aplicação clínica desse hormônio requer padronização adequada de valores de referência, de acordo com o imunoensaio utilizado. Os objetivos deste estudo foram: validar o imunoensaio AMH Gen II (Beckman Coulter Company, TX, USA), estabelecer valores normais de AMH em homens e mulheres saudáveis, avaliar a influência do uso de contraceptivos hormonais, tabagismo e índice de massa corpórea (IMC) sobre os valores de AMH e verificar as concentrações séricas desse hormônio em pacientes com síndrome de Turner (ST), síndrome dos ovários policísticos (SOP) e em meninos com criptorquidismo submetidos ao teste de estímulo com rhCG (gonadotrofina coriônica humana recombinante). A validação do ensaio AMH Gen II foi realizada utilizando protocolo simplificado, conforme as recomendações do Clinical and Laboratory Standards Institute. Para estabelecer valores normais de AMH, 133 mulheres e 135 homens saudáveis foram selecionados prospectivamente. Além disso, 66 pacientes com ST, 29 com SOP e 31 com criptorquidismo foram estudados. A sensibilidade analítica e funcional do ensaio AMH Gen II foram 0,02 e 0,2 ng/mL, respectivamente. Os coeficientes de variação intra e interensaio variaram entre 5,2% - 9,0% e 4,6% - 7,8% em diferentes concentrações, respectivamente. A linearidade, paralelismo e estabilidade das amostras apresentaram percentual de recuperação entre 80% - 120%. O ensaio AMH Gen II correlacionou-se fortemente com o ensaio Immunotech anteriormente utilizado (r = 0,9; p < 0,001). No sexo feminino, os valores séricos de AMH demonstraram declínio progressivo com o aumento da idade (r = -0,4; p < 0,001). Por outro lado, os valores de AMH não diferiram entre usuárias e não usuárias de contraceptivos hormonais, entre tabagistas e não tabagistas, bem como entre mulheres obesas e não obesas. No sexo masculino, a idade não influenciou as concentrações séricas de AMH; no entanto, os valores de AMH foram significativamente reduzidos com o aumento do IMC (r = -0,3, p = 0,008), contudo, esses valores permaneceram dentro do intervalo observado no grupo de indivíduos com peso normal. Todas as pacientes com ST apresentaram valores indetectáveis de AMH. Em contraste, na SOP as concentrações de AMH foram significativamente maiores do que nos controles femininos, mas com 76% de sobreposição entre os dois grupos. Em meninos com criptorquidismo, a concordância entre os valores basais de AMH e a testosterona pósestímulo com rhCG foi de 74%, houve correlação positiva significativa entre os valores basais de AMH e a testosterona pós-estímulo (r = 0,5; p < 0,001). Em conclusão, o ensaio AMH Gen II é confiável para a determinação sérica do AMH. Valores normativos de AMH apresentaram ampla faixa de normalidade em ambos os sexos. Valores reduzidos de AMH em homens obesos encontraram-se dentro da faixa de indivíduos normais. Na ST, o potencial uso do AMH para avaliar a reserva ovariana exige estudos longitudinais. A utilização do AMH como um critério diagnóstico da SOP deve ser associada aos critérios já estabelecidos. Em meninos com criptorquidismo, um AMH normal provê informação útil sobre a função testicular, mas não exclui a necessidade do teste de estímulo com rhCG e pacientes com resultados discordantes necessitam de acompanhamento clínico / The anti-Müllerian hormone (AMH) is a marker of ovarian reserve and testicular function. The clinical application of this hormone requires proper standardization of reference values according to the immunoassay used. The aims of this study were: to validate the AMH Gen II immunoassay (Beckman Coulter Company, TX, USA), to establish reference AMH values in healthy men and women, the influence of hormonal contraceptive use, smoking and body mass index (BMI) on the values of AMH and to check serum concentrations of this hormone in patients with Turner syndrome (TS), polycystic ovary syndrome (PCOS) and in boys with cryptorchidism underwent stimulation test rhCG (recombinant human chorionic gonadotropin).The validation of the AMH Gen II assay was performed using simplified protocol following the recommendations of the Clinical and Laboratory Standards Institute. To establish reference AMH values, 133 healthy women and 135 men were prospectively selected. In addition, 66 patients with TS, 29 with PCOS and 31 with cryptorchidism were studied. The analytical and functional sensitivity of the AMH Gen II assay was 0.02 and 0.2 ng / mL, respectively. Intra and inter-assay coefficients of variation in different concentrations ranged between 5.2-9.0% and 4.6-7.8%, respectively. The linearity, parallelism and stability studies showed recovery % between 80 and 120%. The AMH Gen II assay strongly correlated with the previously used Immunotech assay (r = 0.9, p < 0.001). In females, serum AMH showed a progressive decline with increasing age (r = -0.4, p < 0.001). On the other hand, AMH values did not differ between users and nonusers of hormonal contraceptives, smokers and nonsmokers, obese and non-obese. In males, age did not influence the levels of serum AMH, however, AMH values were significantly reduced with increasing BMI (r = -0.3, p = 0.008), but, these values were within the range observed in the group of subjects with body weight. All TS patients showed undetectable AMH levels. In contrast, in PCOS AMH values were significantly higher than in women controls, but with overlap of 76% between the two groups. In boys with cryptorchidism, the concordance between baseline AMH and testosterone after stimulation with rhCG was 74%, there was a significant positive correlation between baseline AMH values and testosterone values after stimulation (r = 0.5, p < 0.001). In conclusion, AMH Gen II assay is reliable for determining the serum AMH. Reference AMH values showed a wide range in both sexes. Reduced values of AMH in obese men were within the range of normal individuals. In ST, the potential use of AMH to assess ovarian reserve requires longitudinal studies. The use of AMH as a diagnostic criterion of SOP must be based on the association with the criteria already established. In boys with cryptorchidism, a normal AMH provides useful information on testicular function, but does not exclude the need for stimulation test rhCG, patients with discordant results require follow-up

O hormônio anti-Mülleriano (AMH) é um marcador de reserva ovariana e função testicular. A aplicação clínica desse hormônio requer padronização adequada de valores de referência, de acordo com o imunoensaio utilizado. Os objetivos deste estudo foram: validar o imunoensaio AMH Gen II (Beckman Coulter Company, TX, USA), estabelecer valores normais de AMH em homens e mulheres saudáveis, avaliar a influência do uso de contraceptivos hormonais, tabagismo e índice de massa corpórea (IMC) sobre os valores de AMH e verificar as concentrações séricas desse hormônio em pacientes com síndrome de Turner (ST), síndrome dos ovários policísticos (SOP) e em meninos com criptorquidismo submetidos ao teste de estímulo com rhCG (gonadotrofina coriônica humana recombinante). A validação do ensaio AMH Gen II foi realizada utilizando protocolo simplificado, conforme as recomendações do Clinical and Laboratory Standards Institute. Para estabelecer valores normais de AMH, 133 mulheres e 135 homens saudáveis foram selecionados prospectivamente. Além disso, 66 pacientes com ST, 29 com SOP e 31 com criptorquidismo foram estudados. A sensibilidade analítica e funcional do ensaio AMH Gen II foram 0,02 e 0,2 ng/mL, respectivamente. Os coeficientes de variação intra e interensaio variaram entre 5,2% - 9,0% e 4,6% - 7,8% em diferentes concentrações, respectivamente. A linearidade, paralelismo e estabilidade das amostras apresentaram percentual de recuperação entre 80% - 120%. O ensaio AMH Gen II correlacionou-se fortemente com o ensaio Immunotech anteriormente utilizado (r = 0,9; p < 0,001). No sexo feminino, os valores séricos de AMH demonstraram declínio progressivo com o aumento da idade (r = -0,4; p < 0,001). Por outro lado, os valores de AMH não diferiram entre usuárias e não usuárias de contraceptivos hormonais, entre tabagistas e não tabagistas, bem como entre mulheres obesas e não obesas. No sexo masculino, a idade não influenciou as concentrações séricas de AMH; no entanto, os valores de AMH foram significativamente reduzidos com o aumento do IMC (r = -0,3, p = 0,008), contudo, esses valores permaneceram dentro do intervalo observado no grupo de indivíduos com peso normal. Todas as pacientes com ST apresentaram valores indetectáveis de AMH. Em contraste, na SOP as concentrações de AMH foram significativamente maiores do que nos controles femininos, mas com 76% de sobreposição entre os dois grupos. Em meninos com criptorquidismo, a concordância entre os valores basais de AMH e a testosterona pósestímulo com rhCG foi de 74%, houve correlação positiva significativa entre os valores basais de AMH e a testosterona pós-estímulo (r = 0,5; p < 0,001). Em conclusão, o ensaio AMH Gen II é confiável para a determinação sérica do AMH. Valores normativos de AMH apresentaram ampla faixa de normalidade em ambos os sexos. Valores reduzidos de AMH em homens obesos encontraram-se dentro da faixa de indivíduos normais. Na ST, o potencial uso do AMH para avaliar a reserva ovariana exige estudos longitudinais. A utilização do AMH como um critério diagnóstico da SOP deve ser associada aos critérios já estabelecidos. Em meninos com criptorquidismo, um AMH normal provê informação útil sobre a função testicular, mas não exclui a necessidade do teste de estímulo com rhCG e pacientes com resultados discordantes necessitam de acompanhamento clínico / The anti-Müllerian hormone (AMH) is a marker of ovarian reserve and testicular function. The clinical application of this hormone requires proper standardization of reference values according to the immunoassay used. The aims of this study were: to validate the AMH Gen II immunoassay (Beckman Coulter Company, TX, USA), to establish reference AMH values in healthy men and women, the influence of hormonal contraceptive use, smoking and body mass index (BMI) on the values of AMH and to check serum concentrations of this hormone in patients with Turner syndrome (TS), polycystic ovary syndrome (PCOS) and in boys with cryptorchidism underwent stimulation test rhCG (recombinant human chorionic gonadotropin).The validation of the AMH Gen II assay was performed using simplified protocol following the recommendations of the Clinical and Laboratory Standards Institute. To establish reference AMH values, 133 healthy women and 135 men were prospectively selected. In addition, 66 patients with TS, 29 with PCOS and 31 with cryptorchidism were studied. The analytical and functional sensitivity of the AMH Gen II assay was 0.02 and 0.2 ng / mL, respectively. Intra and inter-assay coefficients of variation in different concentrations ranged between 5.2-9.0% and 4.6-7.8%, respectively. The linearity, parallelism and stability studies showed recovery % between 80 and 120%. The AMH Gen II assay strongly correlated with the previously used Immunotech assay (r = 0.9, p < 0.001). In females, serum AMH showed a progressive decline with increasing age (r = -0.4, p < 0.001). On the other hand, AMH values did not differ between users and nonusers of hormonal contraceptives, smokers and nonsmokers, obese and non-obese. In males, age did not influence the levels of serum AMH, however, AMH values were significantly reduced with increasing BMI (r = -0.3, p = 0.008), but, these values were within the range observed in the group of subjects with body weight. All TS patients showed undetectable AMH levels. In contrast, in PCOS AMH values were significantly higher than in women controls, but with overlap of 76% between the two groups. In boys with cryptorchidism, the concordance between baseline AMH and testosterone after stimulation with rhCG was 74%, there was a significant positive correlation between baseline AMH values and testosterone values after stimulation (r = 0.5, p < 0.001). In conclusion, AMH Gen II assay is reliable for determining the serum AMH. Reference AMH values showed a wide range in both sexes. Reduced values of AMH in obese men were within the range of normal individuals. In ST, the potential use of AMH to assess ovarian reserve requires longitudinal studies. The use of AMH as a diagnostic criterion of SOP must be based on the association with the criteria already established. In boys with cryptorchidism, a normal AMH provides useful information on testicular function, but does not exclude the need for stimulation test rhCG, patients with discordant results require follow-up

<p>Nespu&scaron;teni testis predstavlja odsustvo testisa u skrotumu sa jedne ili obe strane. Faktori rizika za pojavu nespu&scaron;tenog testisa obuhvataju genetsku predispoziciju, prevremeno rođenje, nisku porođajnu masu i prenatalnu izloženost endokrinim disruptorima ili duvanskom dimu. Endokrini disruptori se defini&scaron;u kao egzogene supstance koje imaju uticaj na homeostazu organizma i proizvodnju reproduktivnih hormona. U ovoj grupi nalaze se organofosforni pesticidi koji se &scaron;iroko upotrebljavaju u poljoprivredi. Većina organofosfornih pesticida ima antiandrogeni uticaj i uz činjenicu da živimo u pretežno agrarnoj sredini predmet su na&scaron;eg interesovanja. Cilj istraživanja je da se utvrdi razlika u izloženosti organofosfornim pesticidima kori&scaron;ćenjem upitnika kreiranog po modelu standardizovanog Evropskog upitnika QLK 4-1999-01422 kod osoba koje su rodile zdravu mu&scaron;ku decu i osoba koje su rodile decu sa nespu&scaron;tenim testisom. Pored toga, cilj istraživanja je i da se odredi i uporedi vrednost metabolita organofosfornih pesticida (dimetilfosfat, dimetilditiofosfat, dietilfosfat, dietiltiofosfat i dietilditiofosfat) u urinu majki koje su rodile mu&scaron;ku decu sa nespu&scaron;tenim testisima i majki koje su rodile zdravu mu&scaron;ku decu. Metodologija: Rad je radomizovano, prospektivno, kliničko istraživanje sprovedeno na Klinici za ginekologiju i aku&scaron;erstvo Kliničkog centra Vojvodine i Katedri za farmakologiju i toksikologiju Medicinskog fakulteta, Univerziteta u Novom Sadu. U kliničko istraživanje uključeno je 50 porodilja koje su rodile mu&scaron;ku decu sa nespu&scaron;tenim testisima (eksperimentalna grupa) i 53 porodilje koje su rodile zdravu mu&scaron;ku decu (kontrolna grupa) u periodu od oktobra 2012. godine do aprila 2018. godine. Tokom boravka u porodili&scaron;tu ispitanice su popunjavale upitnik o navikama nakon čega im je uzet uzorak urina radi analiziranja nivoa metabolita OF pesticida. Uzorci urina su pripremljeni metodom koju su opisali Wu i saradnici 2010. godine, a potom analizirani na gasnom hromatografu masenom spektrofotometaru marke Agilent 7890A. Rezultati: Ispitivane grupe se ne razlikuju po starosti ispitanica (prosečna starost kontrolne grupe 29,41 &plusmn; 5,58 godina, a eksperimentalne 30,54 &plusmn; 4,87 godina). U obe grupe prosečno je ispitanicama ovo bila druga trudnoća. Ispitanice se nisu razlikovale ni po načinu porođaja. Prosečna gestacijska nedelja trudnoće na porođaju iznosila je 39,45 &plusmn; 1,38 nedelja za kontrolnu grupu i 39,20 &plusmn; 1,38 nedelja za eksperimentalnu grupu, a porođajna masa novorođenčeta 3527,30 &plusmn; 470,16 g u kontrolnoj grupi i 3404,37 &plusmn; 508,20 g u eksperimentalnoj grupi. Statistički značajna razlika postoji u odnosu na mesto stanovanja (50,9 % ispitanica kontrolne grupe i 77,6 % ispitanica eksperimentalne grupe žive u gradu), jedinicu stanovanja (67,9 % kontrolne i 45,7 % ispitanica eksperimentalne grupe žive u kući) i načinu začeća (6 % ispitanica eksperimentalne i 1,9 % ispitanica kontrolne grupe prijavilo je IVF kao način začeća). Skoro polovina ispitanica obe grupe su pu&scaron;ači, a njih 32,7 % kontrolne grupe i 38,8 % eksperimentalne pu&scaron;ile su i tokom trudnoće. Izloženost pesticidima prijavilo je 50,9 % ispitanica kontrolne i 44 % ispitanica eksperimentalne grupe, a profesionalnu izloženost prijavilo je 3 ispitanice kontrolne i 2 ispitanice eksperimentalne grupe. Ispitanice se ne razlikuju ni po poreklu voća i povrća koje konzumiraju, kao ni po vrsti voća koje su konzumirale tokom trudnoće. Prosečne izmerene vrednosti DMF u kontrolnoj grupi iznose 5,604 &plusmn; 6,103 ug/L, a u eksperimentalnoj 4,815 &plusmn; 6,729 ug/L. Izmerene vrednosti DEF u kontrolnoj grupi su 0,408 &plusmn; 0,447 ug/L, a u eksperimentalnoj 0,461 &plusmn; 0,593 ug/L. Nivo DMDTF u kontrolnoj grupi bio je 0,431 &plusmn; 0,508 ug/L, a u eksperimentalnoj 0,547 &plusmn; 0570 ug/L, a DETF 0,403 &plusmn; 0,606 ug/L u kontrolnoj i 0,529 &plusmn; 0,725 ug/L u eksperimentalnoj grupi. Ni jedan metabolit ne pokazuje statistički značajnu razliku u ispitivanim grupama. Slične vrednosti dobijene su i za vrednosti korigovane za nivo kreatinina. Univarijantna regresiona analiza pokazala je da ispitanice koje žive u gradu imaju 3,3 puta veće &scaron;anse da rode dete sa nespu&scaron;tenim testisom, a one koje žive u stanu imaju 2,5 puta veće &scaron;anse za isti ishod. Statistički značajna razlika primećena je u nivou DEDTF u zavisnosti od starosti ispitanica i jedinici stanovanja. Vi&scaron;e vrednosti DETF dobijene su kod ispitanica koje su bile na hormonskoj terapiji tokom trudnoće. Ispitanice koje su prijavile da su bile izložene pesticidima tokom trudnoće u urinu su imale statistički značajno vi&scaron;e vrednosti DMDTF u odnosu na ispitanice koje su se izjasnile da nisu bile izložene pesticidima. Slični rezultati za vrednost DEDTF dobijeni su kod ispitanica koje su se izjasnile da poseduju kućne ljubimce. Statistički vi&scaron;e vrednosti DEF i DETF korigovano za nivo kreatinina dobijene su kod ispitanica koje nisu konzumirale jabuke, a vi&scaron;e vrednosti DEF i DEDTF dobijene su kod ispitanica koje su konzumirale maline i kupine tokom trudnoće. Ostale grupe nisu pokazale statistički značajnu razliku među ispitivanim grupama. Zaključci: Izloženosti trudnica OF pesticidima nije značajno veća u grupi majki koje su rodile decu sa nespu&scaron;tenim testisom u odnosu na izloženost OF pesticidima kod majki zdrave mu&scaron;ke dece. Vrednosti metabolita OF pesticida (dimetilfosfat, dimetilditiofosfat, dietilfosfat, dietiltiofosfat, dietilditiofosfat) u urinu majki koje su rodile mu&scaron;ku decu sa nespu&scaron;tenim testisima nije vi&scaron;a u odnosu na vrednosti metabolita OF pesticida (dimetilfosfat, dimetilditiofosfat, dietilfosfat, dietiltiofosfat, dietilditiofosfat) izmerene u jutarnjem urinu majki koje su rodile zdravu mu&scaron;ku decu.</p> / <p>Undescended testis is the absence of testis in the scrotum on one or both sides. Risk factors for the occurrence of undescended testis include genetic predisposition, premature birth, low birth weight and prenatal exposure to endocrine disruptors or tobacco smoke. Endocrine disruptors are defined as exogenous substances that can affect homeostasis of the organism and the production of reproductive hormones. In this group are organophosphorus pesticides that are widely used in agriculture. Most of organophosphorus pesticides have anti-androgenic effect and with the fact that we live in a predominantly agricultural area, they are the focus of our interest. The aim of the research: The aim of this study is to determine the difference in exposure to organophosphorous pesticides using questionnaires created by standardized European model questionnaire QLK 4-1999-01422 in individuals who gave birth to a healthy male children and women who gave birth to children with undescended testis. In addition, the aim of this study is to determine and compare the value of metabolites of organophosphorus pesticides (dimethylphosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate and diethyldithiophosphate) in the urine of mothers who gave birth to male children with undescended testis and mothers who gave birth to healthy male children. Methodology: This work is randomized, prospective, clinical research conducted at the Clinic for gynaecology and obstetrics of the Clinical center of Vojvodina and the Department of Pharmacology and toxicology of the Faculty of Medicine, University of Novi Sad. This clinical research includes 50 new mothers that gave birth to male children with undescended testes (experimental group) and 53 new mothers that gave birth to healthy male children (control group) in the period from October 2012 to April 2018. During their stay at the maternity hospital the subjects were asked to fill out a questionnaire about habits and to give a urine sample for analyzing the level of metabolites of organophosphus pesticides. Urine samples were then prepared using the method described by Wu and associates 2010, and analyzed on gas chromatograph with a mass spectrophotometer Agilent 7890A brand. Results: Study groups do not differ according to the age of women (average age of control group is 29.41 &plusmn; 5.58 years, and experimental 30.54 &plusmn; 4.87 years). In both groups this was second pregnancy on average. The subjects did not distinguish either by the way of delivery. The average gestational weeks of pregnancy to childbirth was 39.45 &plusmn; 1.38 weeks for the control group and 39.20 &plusmn; 1.38 weeks for the experimental group, and birth weight of newborn was 3527.30 &plusmn; 470.16 g in control group and 3404.37 &plusmn; 508.20 g in the experimental group. There is no statistically significant difference in relation to the place of residence (50.9 % of the control group and 77.6 % of experimental live in the city), the living unit (67.9 % and 45.7 % of the control and experimental groups are living in the house) and the way of conception (6 % of experimental and 1.9 % of the control group reported IVF as a way of conception). Nearly half of both groups are smokers, and 32.7 % of women in the control group and 38.8 % in experimental smoked during pregnancy. Exposure to pesticides reported 50.9 % of mothers in control and 44 % of mothers in the experimental group. Professional exposure was reported by 3 control subjects and 2 subjects in experimental group. The subjects did not differ according to the origin of fruits and vegetables they were consuming, neither regarding the type of fruits they consumed during pregnancy. Average level of dimethylphosphate in control group was 5.604 &plusmn; 6.103 ug/L, and in experimental 4.815 &plusmn; 6.729 ug/l. Levels of diethylphosphate in control group were 0.408 &plusmn; 0.447 ug/L, and in experimental 0.461 &plusmn; 0.593 ug/l. DMDTP level in the control group was 0.431 &plusmn; 0.508 ug/L, and in experimental 0.547 &plusmn; 0570 ug/L, and the DETP was measured 0.403 &plusmn; 0.606 ug/L in control, and 0.725 &plusmn; 0.529 ug/L in the experimental group. These metabolites showed no statistically significant difference in the examined groups. Similar values are obtained for the adjusted values for creatinine level. Univariate regression analysis showed that the subjects who live in town are 3.3 times more likely to have child with undescended testis, and those who live in the apartment are 2.5 times more likely for the same outcome. Statistically significant difference was noticed in DEDTP level depending on the age of the subject and the living unit. Higher levels of DETP metabolites were detected in subjects that have been on hormonal therapy during pregnancy. The subjects who reported being exposed to pesticides during pregnancy had statistically significantly higher DMDTP values in relation to the subjects that were not exposed to pesticides. Similar results are obtained for the DEDTP level with higher levels in subjects owning pets. Statistically higher levels of DEP and DETP adjusted for creatinine were obtained in subjects that were not reporting eating apples, and higher levels of DEP and DEDTP were obtained in subjects that consumed raspberries and blackberries during pregnancy. Other groups showed no statistically significant difference between the study groups. Conclusion: Exposure of pregnant women to OP pesticides is not significantly greater in the group of mothers who gave birth to children with undescended testis in relation to exposure to OP pesticides in mothers of healthy male children. The level of OP metabolites (dimethylphosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate and diethyldithiophosphate) in the urine of mothers who gave birth to children with undescended testis is not higher in relation to the levels of OP metabolites (dimethylphosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate and diethyldithiophosphate) recorded in urine of mothers who gave birth to healthy male children.</p>