Recueil chirurgical de spermatozoïdes dans un groupe de 50 patients avec un antécédent de cryptorchidie bilatérale: facteurs pronostics d’extraction

Bilateral cryptorchidism is a severe factor of male infertility. We evaluated the results of testicular sperm extraction in 50 men with nonobstructive azoospermia associated with cryptorchidism between 1995 to 2001. We evaluated clinical parameters predictive of successful sperm extraction. Serum follicle stimulating hormone (FSH) was elevated in 67% of cases and testicular volume was decreased in 70% of cases, confirming the secretory origin of the azoospermia. Serum follicle stimulating hormone (FSH), testicular volume and histological parameters were examined as predictive factors for sperm recovery. The positive sperm recovery rate was 68%. As in the population of men with nonobstructive azoospermia, the sperm recovery rate for patients with a history of orchidopexy is approximately 68% and there are currently no clinical parameters predicting successful sperm retrieval in this subpopulation of patients.     

Sequence diversity of MHC class-II <Emphasis Type="BoldItalic">DRB</Emphasis> gene in gazelles (<Emphasis Type="BoldItalic">Gazella subgutturosa</Emphasis>) raised in Sanliurfa of Turkey

In this study, we aimed to assess the sequence diversity of major histocompatibility complex (MHC) class-II DRB gene at exon 2 in gazelles raised in Sanliurfa Province of Turkey. Twenty DNA samples isolated from gazelles (Gazella subgutturosa) were used for sequencing exon 2 of MHC class-II DRB gene. Target region was amplified by polymerase chain reaction (PCR) and their products were directly sequenced. Nine of these 20 samples yielded unambiguously readable sequences. Three of the nine samples were homozygotes and each showed different sequences. A 262-bp sequence obtained from the three homozygote samples were submitted to GenBank (accession numbers: KC309405, KC309406 and KC309407). Using an allele specific PCR, we detected 10 additional haplotypes. Among 13 haplotypes, 45 nucleotide positions were polymorphic and most of the polymorphic nucleotide positions localized at peptide-binding region (PBR). Rates of nonsynonymous substitutions were significantly higher than synonymous substitutions at PBR. Phylogenetic analysis of the haplotypes showed that 10 haplotypes of the gazelles were clustered together while three were clustered with ovine and bovine haplotypes. The results indicated that at least 13 haplotypes at exon 2 of MHC class-II DRB gene were showing high degree of nucleotide and amino acid diversity, and certain haplotypes of G. subgutturosa were more similar to haplotypes from sheep or cattle than to each other. Rates of synonymous and nonsynonymous substitutions suggested that positive selection was a driving force for diversity at this locus in G. subgutturosa.     

Exploration du plasma séminal humain: approche physico-chimique

Human seminal plasma glycerylphosphorylcholine (GPC), choline, citrate, and lactate were analysed by measuring the peak area of 1H nuclear magnetic resonance spectra (1H-MRS) in patients with spermatogenic failure or and obstructive azoospermia. The peak area ratios choline/citrate as well as choline/lactate were significantly different (p<0.01) between spermatogenic failure and obstructive azoospermia groups. When the serum FSH values was normal in spermatogenic failure men and obstructive azoospermia, a significant difference, was found in the GPC/choline ratio (P<0.001). The GPC/choline ratio appears to be a very important parameter not only to differentiate between spermatogenic failure and obstructive azoospermia when the FSH values are normal but also between differents forms of spermatogenic failure men. These results demonstrate the potential use of 1H-MRS on human seminal plasma in male infertility management.     

Immunofluorescent synaptonemal complex analysis in azoospermic men

The molecular cause of germ cell meiotic defects in azoospermic men is rarely known. During meiotic prophase I, a proteinaceous structure called the synaptonemal complex (SC) appears along the pairing axis of homologous chromosomes and meiotic recombination takes place. Newly-developed immunofluorescence techniques for SC proteins (SCP1 and SCP3) and for a DNA mismatch repair protein (MLH1) present in late recombination nodules allow simultaneous analysis of synapsis, and of meiotic recombination, during the first meiotic prophase in spermatocytes. This immunofluorescent SC analysis enables accurate meiotic prophase substaging and the identification of asynaptic pachytene spermatocytes. Spermatogenic defects were examined in azoospermic men using immunofluorescent SC and MLH1 analysis. Five males with obstructive azoospermia, 18 males with nonobstructive azoospermia and 11 control males with normal spermatogenesis were recruited for the study. In males with obstructive azoospermia, the fidelity of chromosome pairing (determined by the percentage of cells with gaps [discontinuities]/splits [unpaired chromosome regions] in the SCs, and nonexchange SCs [bivalents with 0 MLH1 foci]) was similar to those in normal males. The recombination frequencies (determined by the mean number of MLH1 foci per cell at the pachytene stage) were significantly reduced in obstructive azoospermia compared to that in controls. In men with nonobstructive azoospermia, a marked heterogeneity in spermatogenesis was found: 45% had a complete absence of meiotic cells; 5% had germ cells arrested at the zygotene stage of meiotic prophase; the rest had impaired fidelity of chromosome synapsis and significantly reduced recombination in pachytene. In addition, significantly more cells were in the leptotene and zygotene meiotic prophase stages in nonobstructive azoospermic patients, compared to controls. Defects in chromosome pairing and decreased recombination during meiotic prophase may have led to spermatogenesis arrest and contributed in part to this unexplained infertility.     

Polymorphism of the serotonin 2A receptor gene (5HTR2A) and personality traits

The interindividual variation of temperament features (such as anxiety, neuroticism, harm avoidance) is determined, in particular, by allele polymorphism of genes involved in serotonin metabolism and has earlier been associated with the insertion/deletion polymorphism of the serotonin transporter gene. Polymorphic alleles of the serotonin 2A receptor gene (5HTR2A) were tested for association with personality traits assessed with several tests. The T102C and A1438G polymorphisms were associated with a variation in emotionality, activity, and sociability, which are integral characteristics of temperament. With each polymorphism, differences were significant only between heterozygotes and homozygotes. Carriers of T102C genotype A1/A2 displayed a lower level of anxiety-related traits, a higher score on scale Hypomania, and a lower score on scale Social Introversion and were assumed to have higher activity and sociability. Carriers of A1428G genotype A/G differed from homozygotes G/G in having a lower level of social introversion and a lower score on scale No close friends, which testified to higher sociability of heterozygotes. Thus, the polymorphic alleles of SHTR2A proved to be associated with personality traits in mentally healthy people.     

Variation in particular biochemical indicators,cytokine and adipokine profiles of the blood,and the structural and functional parameters of the liver in patients with nonalcoholic fatty liver disease and different genotypes by the polymorphic locus A313G of the <Emphasis Type="Italic">GSTP1</Emphasis> gene

Variation in particular biochemical indicators, cytokine and adipokine profile parameters of the blood, and the structural and functional parameters of the liver have been studied in patients with nonalcoholic fatty liver disease (NAFLD) and different genotypes by the polymorphic locus A313G of GSTP gene (rs 1695). It has been established that the G allele of the GSTP gene (A313G) is significantly more frequently met in NAFLD patients than in healthy individuals. Higher activity of alanine aminotransferase and higher level of leptin, as well as lower adiponectin blood content, were recorded in the carriers of the G allele of the GSTP gene (A313G) as compared to patients with the given gene AA genotype. Higher interleukin-10 blood content was also observed in homozygous G allele carriers of NAFLD patients compared with the patients with the AA and AG genotypes. The patients with the G allele of the GSTP1 gene had a larger right lobe of liver than homozygous carriers of the given gene A allele.     

Genetic diversity and mating system of <Emphasis Type="Italic">Pinus brutia</Emphasis> var. <Emphasis Type="Italic">stankewiczii</Emphasis> Sukacz. in small localities of Sudak (Crimea)

The comparative analysis of genetic variation at 12 polymorphic isozyme loci, as well as the mating system, has been carried out in mature trees and their seed progenies in three small localities of Pinus brutia var. stankewiczii Sukacz. in the region between the town of Sudak and settlement of Novyi Svet in Crimea. We found that embryos maintain the same allelic diversity as parent trees; however, their observed heterozygosity, on average, is lower by 37.4%. The significant deviation of genotype distribution from the expected ratios caused by the deficiency of heterozygotes was observed at 8 out of 12 loci. The multilocus estimate of the out-crossing rate (t _m ) in populations varied from 68.9 to 94.9% with the average of 80.7%.     

Microsatellite Variability of Pacific Herring <Emphasis Type="Italic">Clupea pallasii</Emphasis> Valenciennes, 1847 from the Sea of Okhotsk and Bering Sea

The genetic variability of ten microsatellite loci was examined in samples of the herring from the Sea of Okhotsk and the Bering Sea. All loci were polymorphic; the expected heterozygosity estimates varied in the range of 0.3–94.3% (mean 66.7%). The degree of genetic differentiation of the herring was statistically significant (θ = 1.38%). The level of pairwise genetic differentiation F_ST was–0.002–0.046; R_ST was–0.003–0.166. Genetic differentiation of the herring from the Sea of Okhotsk and the Bering Sea correlated with the spatial-geographic structure of the species in the studied range on the basis of F_ST (P = 0.001).     

Analysis of molecular genetic study on the polymorphic C-159T locus of the <Emphasis Type="Italic">CD14</Emphasis> gene in children with increased risk of recurrent episodes of acute obstructive bronchitis

A molecular genetic study of the polymorphic C-159T locus of the CD14 gene has been carried out in 31 children with recurrent episodes of acute obstructive bronchitis as well as in a cohort of 50 randomly sampled subjects forming the control group. As a result of the study, no higher statistically significant frequency of the CC, CT, and TT genotypes has been recorded at the polymorphic C-159T locus of the CD14 gene among children of the main group compared with the control. However, there is a trend towards a more frequent observation of the TT genotype among children with recurrent episodes of acute obstructive bronchitis compared with the control group (32.25 vs. 22.0%).     

Polymorphisms of caprine <Emphasis Type="Italic">GnRHR</Emphasis> gene and their association with litter size in West African Dwarf goats

Gonadotropin-releasing hormone receptor (GnRHR) gene is considered a candidate gene for litter size due to its critical role in regulating the activities of hypothalamo-pituitary-gonadal axis which synthesizes and releases gonadotropins. This study was designed to identify mutations within the caprine GnRHR gene and investigate their association with litter size at various parities. Polymorphisms scanning and genotyping of GnRHR gene in West African Dwarf (WAD) goats (n?=?226) revealed three single nucleotide polymorphisms (SNPs), one mutation (g.-29T?>?G) was detected within 5′UTR region while two others (g.48G?>?A and g.209T?>?G) were identified in exon 1. Mutation at g.209T?>?G locus resulted in amino acid change from Methionine to Arginine at position 70 on the polypeptide residue. Based on heterozygosity and polymorphism information content (PIC), WAD goat population diversity at the SNP loci was moderate. Strong linkage disequilibrium (LD) (r²?>?0.98) existed among the detected mutations resulting in three observed haplotypes, two (T-G-T and G-A-G) had cumulative frequency of >?97%. The mutation within 5′UTR region of GnRHR gene (g.-29T?>?G) is novel, being reported in goats for the first time. Association analysis revealed a significant (p?<?0.05) association between allele G at g.-29T?>?G with higher mean litter size for homozygous (GG) mutant does compared with heterozygotes (GT) or homozygotes (TT), while the relationship between SNPs at the two loci detected in exon 1 and litter size was not significant.     

Association of the <Emphasis Type="Italic">POLG1</Emphasis> T(−365)C, <Emphasis Type="Italic">ANT1</Emphasis> G(−25)A,and <Emphasis Type="Italic">PEO1</Emphasis> G(−605)T polymorphisms with diabetic polyneuropathy in patients with Type 1 diabetes mellitus

This study is dedicated to a search for the association of the polymorphic markers T(?365)C of the POLG1 gene G(?25)A of the ANT1 gene and G(?605)T of the PEO1 gene with diabetic polyneuropathy (DPN) in Type 1 diabetes mellitus (DM1) patients. All patients were ethnic Russian Moscow residents, with DM1 records of no more than 5 years and DPN or DM1 records of more than 10 years but without DPN. We found that the polymorphic marker T(?365)C of POLG1 was associated with DPN in Russian patients with DM1. The carriers of the C allele and the CC genotype had a higher risk of DPN development (OR = 1.62; CI = 1.11–2.38; and OR = 1.76; CI = 0.99–3.13; respectively). In contrast, the T allele carrier status and the TT genotype were associated with a lower DPN risk (OR = 0.62, CI = 0.42–0.90; and OR = 0.61; CI = 0.35–1.07; respectively). We found no association of the polymorphic markers G(?25)A of ANT1 or G(?605)T of PEO1 with DPN in Russian DM1 patients living in Moscow.     

Impact of Single Nucleotide Polymorphisms of Base Excision Repair Genes on DNA Damage and Efficiency of DNA Repair in Recurrent Depression Disorder

Elevated level of DNA damage was observed in patients with depression. Furthermore, single nucleotide polymorphisms (SNPs) of base excision repair (BER) genes may modulate the risk of this disease. Therefore, the aim of this study was to delineate the association between DNA damage, DNA repair, the presence of polymorphic variants of BER genes, and occurrence of depression. The study was conducted on peripheral blood mononuclear cells of 43 patients diagnosed with depression and 59 controls without mental disorders. Comet assay was used to assess endogenous (oxidative) DNA damage and efficiency of DNA damage repair (DRE). TaqMan probes were employed to genotype 12 SNPs of BER genes. Endogenous DNA damage was higher in the patients than in the controls, but none of the SNPs affected its levels. DRE was significantly higher in the controls and was modulated by BER SNPs, particularly by c.977C>G–hOGG1, c.972G>C–MUTYH, c.2285T>C–PARP1, c.580C>T–XRCC1, c.1196A>G–XRCC1, c.444T>G–APEX1, c.-468T>G–APEX1, or c.*50C>T–LIG3. Our study suggests that both oxidative stress and disorders in DNA damage repair mechanisms contribute to elevated levels of DNA lesions observed in depression. Lower DRE can be partly attributed to the presence of specific SNP variants.     

Role of genetic variations of <Emphasis Type="Italic">chitinase 3</Emphasis>-<Emphasis Type="Italic">like 1</Emphasis> in bronchial asthmatic patients

Background

Single nucleotide polymorphisms (SNPs) in chitinase 3-like 1 (CHI3L1) are associated with bronchial severity and pulmonary function. CHI3L1 proteins are involved in both innate and adaptive immune responses; however, to date, the correlation of these SNPs and their age of onset of bronchial asthma has not been demonstrated.

Methods

To address the role of these genetic variations, 390 patients with well-controlled bronchial asthma and living in Japan were recruited, genotyped, and had a pulmonary function test performed on them in this study. To analyze the concentration levels of CHI3L1 protein, bronchial lavage fluids were examined.

Results

Forced expiratory volume in one second, %predicted (%FEV1), was significantly decreased in homozygotes of rs1214194 compared to heterozygotes and wild type. The age of onset of adult bronchial asthma was significantly younger in GG homozygotes of rs4950928 and AA homozygotes of rs1214194 than in the other two genotypes. The concentration of CHI3L1 protein in bronchial lavage fluid increased in both homozygotes of rs4950928 and rs1214194.

Conclusions

Our study demonstrated that the homozygotes of rs4950928 and rs1214194 of CHI3L1 might predict an early onset of bronchial asthma and have the propensity to promote airway remodeling.Trial registration JMA-IIA00045 remodeling-ICS

     

Genotypical features in the expression of allozymes of β-specific hydrolase of carboxylic esters in wild-type <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The expression level of electrophoretically separated S- and F-allozymes of β-specific esterase (EC 3.1.1.2) in genotypes of wild-type Drosophila melanogaster (males and females) that are monozygous or heterozygous with respect to the locus β-Est is determined by means of computerized densitometry; α-naphthylacetate, β-naphthylacetate, and α-naphthylpropionate are used as the substrates. The intensity of the expression of the esterase is judged from the quantity of reaction product created as a result of simultaneous azo coupling between naphthol and diazonium in 4, 24, 44, and 64 min incubation times. Reliable differences in the expressions of the S- and F-allozymes as a function of the structure of the β-Est locus of genotypically distinct individuals are established. In all the variant experiments, a higher level of summary activity of the S- and F-allozymes of the β-esterase of the heterozygotes by comparison with the individual activity of the F-and S-allozymes of the corresponding homozygotes was demonstrated, independently of the sex of the Drosophila individual. A comparative estimate of the temporal dynamics of the expression of in vitro allozymes of the dominant homozygotes (β-Est _S /β-Est _S ), heterozygotes (β-Est _S /β-Est _F ), and recessive homozygotes (β-Est _F /β-Est _F ) is performed. Possible mechanisms for the occurrence of heterosis according to the character of expression of S- and F-allozymes of β-esterase on the basis of the theory of biochemical enrichment of heterozygote genotypes are considered.     

Association of rs1057035polymorphism in microRNA biogenesis pathway gene (<Emphasis Type="Italic">DICER1</Emphasis>) with azoospermia among Iranian population

Since genes involved in microRNA biogenesis pathways have a main role in impaired spermatogenesis, in this research, we evaluated different genotypes frequency of seven single-nucleotide polymorphisms in DICER1 and DROSHA genes. Different genotypes frequency of DICER1 (rs12323635, rs1057035, rs13078 and rs3742330) and DROSHA (rs10719, rs642321 and rs2291102) were determined by sequencing method in 385 infertile men and 120 fertile controls. It was found that CC genotype (P?=?0.000) and C allele (P?=?0.0) of rs1057035 T?>?C polymorphism were associated with idiopathic male infertility (azoospermia). Gene expression study in blood and testis samples was done by real time PCR technique. Our results showed significant under expression of DICER1 gene in blood and testis tissues of azoospermic samples (P?<?0.05), but we did not observed significant difference in expression ratio between infertile men with and without C allele of rs1057035 SNP (P?>?0.05). The results of this study showed that among the studied variants, only one of them in DICER1 might be associated with azoospermia, but additional studies needs in different populations and ethnics.     

Isolation and characterization of microsatellite loci from <Emphasis Type="Italic">Spodoptera exigua</Emphasis> (Lepidoptera: Noctuidae)

Ten polymorphic microsatellite loci were isolated and characterized from Spodoptera exigua (Hübner), which is a worldwide economic pest on various crops. The isolated loci were polymorphic, with 3–15 alleles in 40 individuals from four regional populations in Korea. The analyses revealed that 40 individuals had different multilocus genotypes, with the expected heterozygosity ranging from 0.133 to 0.899. Eight of the ten loci did not deviate significantly from Hardy-Weinberg equilibrium. The isolated markers will facilitate population genetic studies of S. exigua.     

Isolation and characterization of 14 microsatellite markers in <Emphasis Type="Italic">Macrodasyceras</Emphasis><Emphasis Type="Italic">hirsutum</Emphasis> (Hymenoptera: Torymidae)

Macrodasyceras hirsutum Kamijo is the seed parasitoid wasp of the bird-dispersed, dioecious tree, Ilex integra Thunb. The wasp reduces the level of dispersal mutualism between the Ilex tree and its frugivorous birds by manipulating the color of mature berries. The female trees do not blossom every year and sometimes change sex. Thus, the reproduction biology of I. integra affects the population size and structure of M. hirsutum in a forest and consequently influences the seed dispersal mutualism between the tree and birds, because of limited ability of adult locomotion. To investigate the wasp population structure with reference to the dispersal mutualism between trees and birds, we isolated 14 microsatellite loci of M. hirsutum wasps. Every locus was polymorphic among 20 females, with 3–13 alleles per locus, without linkage disequilibrium. The observed and expected heterozygosities ranged from 0.100 to 0.900 and 0.099 to 0.818, respectively, indicating their utility in molecular analyses of the wasp population.