Description of Echinococcus granulosus genotypes in human hydatidosis in a region of southern Chile

INTRODUCTION: Echinococcus granulosus species has a wide variety in both geography and hosts; indeed, 10 genotypes have been reported in studies on material of animal origin. The aim of this study was to genotype E. granulosus obtained from human hydatid cysts. MATERIALS AND METHODS: The hydatid fluid and sand was collected from patients who underwent surgery for hepatic and pulmonary hydatidosis at Hospital Regional in Temuco, Chile, between 2004 and 2005. Two PCR systems were used: PCR Eg 9 and PCR Eg 16. The RsaI enzyme was used for RFLP. The genotype was confirmed using the sequence of one fragment of 366 bp from a mitochondrial gene (cox1). RESULTS: The DNA of protoscolices from 24 samples was analyzed, 4 of them from pulmonary cysts and 20 from hepatic cysts. The 366 bp fragment was amplified in 20 out of 24 samples (83.3%). Enzymatic digestion revealed the presence of 3 possible genotypes: in 20 out of 21 samples (95,2%), a restriction was observed corresponding to the G1 or G7 genotypes; in the remaining sample genotype G4 or G7 was observed. Sequencing confirmed the presence of G1 genotype for 19 samples and G6 genotype for the remaining sample (G4 or G7 according to PCR-RFLP). CONCLUSION: The PCR-RFLP technique enabled three possible genotypes present (G1 or G7, G4 or G7) to be established. Sequencing allowed us to decisively identify the G1 and G6 genotypes in our study group. Previous studies agree with the identification of the G1 genotype in our country. We consider it significant that the G6 genotype is present in Chile for its epidemiological implications.     

Molecular evidence for the presence of a G7 genotype of Echinococcus granulosus in Slovakia

Variability in Echinococcus granulosus is very important epidemiologically since strain characteristics may influence local patterns of transmission of hydatid disease. To classify the genotype presented in pig protoscoleces of the Slovak territory, a DNA-based approach has been used. Nucleotide sequences for a 471 bp region of the mitochondrial NADH dehydrogenase 1 (ND1) gene revealed a substantial affinity of isolates examined to the G7 genotype. Only a 0.9-3.4% sequence variation was recorded for E. granulosus samples compared with the reference G7 variant. To distinguish between G7 and G9 genotypes not differing in ND1 sequences, isolates were additionally examined by PCR-RFLP analysis of the nuclear ITS1 region. The resulting two-banded pattern is characteristic for the G7 strain. The data presented thus provides the first explicit evidence of the G7 genotype in the Slovak region.     

Unoccupied prolactin binding components of the benign and malignant human prostate in a subclinical and clinical procedure

Optimal conditions for the quantitation of free prolactin binding components of human prostatic tissue obtained by TURP were studied by applying γ receptor assay. The radioligand used was ¹²⁵I-prolactin. Significantly greater heat stability of the prostate membrane prolactin binding sites, when compared to that of androgen cytoplasmic receptors, was confirmed. The saturability and specificity of the prolactin binding components was demonstrated by the results of both Scatchard plot analysis and displacement studies. Free prolactin receptors were found in none of the poorly differentiated (G3) prostatic tumors examined, and only in 62.5% of medium differentiated (G2) prostatic malignancies. The majority of tissue specimens coming from patients with either BPH or well differentiated prostatic tumor (G1) contain measurable amounts of free prolactin membrane binding components. In the present study we report also the case in which the change in tumor differentiation toward a higher grade (G2 to G1, provoked by the successful chemohormonal treatment) is accompanied with the appearance of previously absent free prolactin binding components. In histologically proven BPH tissue specimens free prolactin receptor negative status has been found in most patients with a slight increase in serum PAP values, while receptor rich status was detected in the majority of those with elevated PSA concentrations. We believe therefore that the prolactin receptor values, when used as part of the multivariable analysis, may participate in further delineation of the role of prolactin in the development of prostate cancer, but may also play a role in a subclinical prediction related to the conversion of either an adenoma or a latent adenocarcinoma to the clinically manifest prostatic malignancy.     

Association of a -1997G-->T polymorphism of the collagen Ialpha1 gene with bone mineral density in postmenopausal Japanese women

Genetic variants that affect collagen Ialpha1 metabolism may be important in the development of osteoporosis or osteoporotic fractures. A -1997G-->T polymorphism in the promoter of the collagen Ialpha1 gene (COL1A1) was shown to be associated with bone mineral density (BMD) for the lumbar spine in postmenopausal Spanish women. The relation of this polymorphism to BMD in Japanese women or men has now been examined in a population-based study. The subjects (1,110 women, 1,126 men) were 40 to 79 years of age and were randomly recruited for a population-based prospective cohort study of aging and age-related diseases. BMD for the lumbar spine, right femoral neck, right trochanter, and right Ward's triangle was measured using dual-energy x-ray absorptiometry. Genotypes for the -1997G-->T polymorphism of COL1A1 were determined with a fluorescence-based allele-specific DNA primer assay system. When all women were analyzed together, BMD for the lumbar spine and trochanter was significantly lower in subjects with the COL1A1 *G/*G genotype than in those in the combined group of COL1A1 *G/*T and COL1A1 *T/*T genotypes. When postmenopausal women were analyzed separately, BMD for the femoral neck and trochanter was also significantly lower in those with the COL1A1 *G/*G genotype than in those with the COL1A1 *G/*T genotype or those in the combined group of COL1A1*G/*T and COL1A1 *T/*T genotypes. BMD was not associated with -1997G-->T genotype in premenopausal women or in men. Multivariate regression analysis revealed that -1997G-->T genotype affected BMD at various sites with a variance of 0.46-0.62% for all women and 0.61-1.01% for postmenopausal women. The -1997G-->T genotype was not related to the serum concentration of osteocalcin, the serum activity of bone-specific alkaline phosphatase, or the urinary excretion of deoxypyridinoline or cross-linked N-telopeptides of type I collagen in men or in premenopausal or postmenopausal women. These results suggest that COL1A1 is a susceptibility locus for reduced BMD in postmenopausal Japanese women.     

Evaluation of genetic behavior of some Egyption Cotton genotypes for tolerance to water stress conditions

Water stress is a critical abiotic stress for plant reduction in arid and semiarid zones and, has been discovered to be detrimental to the development of seedlings as well as the growth and physiological characteristics of many crops such as cotton. The objectives of our study were to determine the combining ability and genetic components for five quantitative traits [(leaf area (LA), leaf dry weight (LDW), plant height (PH), fiber length (2.5 percent SL), and lint cotton yield/plant (LCY/P)] under water shortage stress, a half diallel cross between six cotton genotypes representing a wide range of cotton characteristics was evaluated in RCBD with four replications. The genotype mean squares were significant for all traits studied, demonstrating significant variation among genotypes for all characters under water shortage stress. LCY/P had the highest phenotypic and genotypic correlation co-efficient with PH, LDW, and LA shortage. The highest direct effect on lint cotton yield was exhibited by leaf area (3.905), and the highest indirect effects of all traits were through LA, with the exception of 2.5 percent SL, which was through LDW. The highest dissimilarity (Euclidean Distance) between parental genotypes was between G.87 and G.94, followed by G.87 and Menoufi. G.94 was also a well-adapted genotype, and the combinations G.87 x G.94 and G.87 x Menoufi may outperform their parents. The combining ability analysis revealed highly significant differences between parental GCA effects and F1 crosses SCA effects. The variation of GCA and SCA demonstrated the assurance of additive and non- additive gene action in the inheritance of all traits studied. In terms of general combining ability (GCA) effects, parental genotype G.94 demonstrated the highest significant and positive GCA effects for all traits studied, with the exception of 2.5 percent SL, where G.87 revealed the highest significant and positive GCA effects. The effects of specific combining ability (SCA) revealed that the cross (G.87 x2G.94) revealed stable, positive, and significant SCA for all of the studied traits.     

Nucleotide sequence polymorphism within exon 4 of the bovine prolactin gene and its associations with milk performance traits

Prolactin plays an important regulatory function in mammary gland development, milk secretion, and expression of milk protein genes. Hence the PRL gene is a potential quantitative trait locus and genetic marker of production traits in dairy cattle. We analysed the sequence of the PRL gene to investigate whether mutations in this sequence might be responsible for quantitative variations in milk yield and composition. Using SSCP and direct sequencing, we detected six single-nucleotide polymorphisms within a 294-bp prolactin gene fragment involving exon 4. All detected mutations were silent with respect to the amino acid sequence of the protein. PCR-RFLP genotyping of SNP 8398 R (RsaI) was used to assess allele frequencies in 186 Black-and-White cows (0.113 and 0.887 for A and G, respectively) and in 138 Jersey cows (0.706 and 0.294 for A and G, respectively). Black-and-White cows with genotype AG showed the highest milk yield, while cows with genotype GG showed the highest fat content.     

Genetic and biochemical diversity of Gardnerella vaginalis strains isolated from women with bacterial vaginosis

Gardnerella vaginalis is considered a substantial player in the progression of bacterial vaginosis (BV). We analysed 17 G. vaginalis strains isolated from the genital tract of women diagnosed with BV to establish a potential link between genotypes/biotypes and the expression of virulence factors, vaginolysin (VLY) and sialidase, which are assumed to play a substantial role in the pathogenesis of BV. Amplified ribosomal DNA restriction analysis revealed two G. vaginalis genotypes. Gardnerella vaginalis isolates of genotype 2 appeared more complex than genotype 1 and were subdivided into three subtypes. Biochemical typing allowed us to distinguish four different biotypes. A great diversity of the level of VLY production among the isolates of G. vaginalis may be related to a different cytotoxicity level of the strains. We did not find any correlation between VLY production level and G. vaginalis genotype/biotype. In contrast, a link between G. vaginalis genotype and sialidase production was established. Our findings on the diversity of VLY expression level in different clinical isolates and linking sialidase activity with the genotype of G. vaginalis could help to evaluate the pathogenic potential of different G. vaginalis strains.     

Candidate genes for milk production traits in Portuguese dairy sheep

In this study, the influence of the β-lactoglobulin, prolactin and α_S1-casein variants on milk production traits of three Portuguese sheep breeds, including Serra da Estrela, White Merino and Black Merino, was analyzed. The genetic variants of each marker were identified using PCR-RFLP. β-Lactoglobulin genotype AA was associated with lower milk yield in Serra da Estrela and Merino ewes. This marker also affected milk fat content in Serra da Estrela and protein content in Merino. First results regarding the influence of the sheep prolactin gene on milk production traits are reported. In the Serra da Estrela breed, ewes carrying prolactin genotype AA were significantly associated with lower milk yield, but this influence of prolactin genotypes on milk yield was not detected in Merino. Prolactin genotypes were also significantly associated with milk fat and protein contents in the Serra da Estrela breed. A suggestive effect of the α_S1-casein locus on milk yield was detected in Serra da Estrela, but no associations were found between the variants of this marker with milk fat and protein content.     

鸽黑素皮质素受体3、4(MC3R、MC4R)基因多态性与生长性状的相关分析

利用PCR-SSCP技术和DNA测序方法检测广东石岐肉鸽和哈尔滨地区灰色家鸽MC3R和MC4R基因部分编码区序列的单核苷酸多态性, 分析了MC3R基因T91G突变位点和MC4R基因A903G突变位点导致的基因型与两群体鸽生长和体组成性状的关系。结果表明, 这两个多态位点所导致的基因型对石岐肉鸽活重、屠体重、全净膛重均有显著影响(P<0.05); 另外, 利用这两个突变位点所产生的合并基因型在鸽群体中与生长和体组成性状作最小二乘分析, 结果表明, 两位点合并后的基因型对全净膛重影响显著(P<0.05)。多重比较结果表明, BBAA型全净膛重显著大于AABB型, BBAA型对于体重增长是有利基因型。     

The peculiarities of polymorphism of XPD and XRCC1 repair genes in cells of Down and Ehlers-Danlo syndrome patients characterized by increased radiosensitivity

Codon 312 and 751 polymorphisms ofXPD gene and codon 399 polymorphism of XRCC1 gene of peripheral blood lymphocytes in patients with Down syndrome (DS) (46 individuals), Ehlers-Danlo syndrome (EDS) (47 individuals) and in a group of healthy donors (control) (40 individuals) have been studied. Frequency of XPD genotype (G312G) coding for the most effectively functioning form of XPD protein was lower in patients with DS (26%) than in a group of healthy donors (42.5%) (p = 0.035), whereas no significant differences with the control were revealed for this codon in patients with EDS. No patients with XPD genotype (C751C) (p = 0.036) were revealed in a group of EDS patients, while this genotype was found in 16% of a group of healthy donors and in 17% of patients with DS. The trend of XRCC1 genotype frequency reduction (A399A) (p = 0.085) in EDS patients (3.9%) compared with the group of healthy donors (13.5%) and DS patients (13.3%) has been obtained. These data show that polymorphisms of the excision repair genes under study are accompanied by an elevated individual radio sensitivity in patients with DS. Genes investigated (their polymorphic variants) did not participate in the mechanisms for radio sensitive phenotype formation in EDS patients.     

Prolactin storage in a clonal strain of rat pituitary tumor cells is cell-cycle dependent

GH4C1 cells (CH cells) are a clonal strain of rat pituitary tumor cells which secrete prolactin. We measured intracellular prolactin at different stages of the cell cycle using flow microfluorometry. Prolactin was stained by an indirect immunocytochemical technique using fluorescein isothiocyanate (FITC)-conjugated antiserum, and DNA was stained simultaneously with propidium iodide. We found that prolactin storage in GH cells was cell-cycle dependent; prolactin storage increased as cells passed from G1 to S to G2 + M. We have shown previously that insulin and 17 beta-estradiol act synergistically to increase intracellular prolactin three- to sevenfold and slow the rate of cell growth to approximately 70% of control cells. In this study we observed that insulin and estradiol increased prolactin storage at each stage of the cell cycle but did not affect the cell-cycle distribution of the population even though cell growth was slowed. We conclude that insulin and estradiol did not increase prolactin storage by affecting the cell-cycle distribution of the population.     

Prolactin receptor as a candidate gene for prolificacy of small tail han sheep

DNA polymorphism of the ovine prolactin receptor gene (PRLR) was investigated and used to study its effect on litter size in sheep. By means of PRLR gene sequence homology between sheep and human, three primer pairs were designed for polymerase chain reaction (PCR) amplification within intron 1 and exon 10 of the PRLR gene in sheep. In these parts of the gene the single nucleotide polymorphisms were detected by PCR-single strand conformation polymorphism (SSCP) in 314 Small Tail Han ewes. These poly-morphisms were used to study the associations with litter size. The results indicated that there were three genotypes (AA, AB and BB) detected by three primer pairs. For three primer pairs the frequency of allele A was 0.96, 0.79, 0.68; and the frequency of allele B was 0.04, 0.21, 0.32, respectively. The frequency of genotype AA was 0.93, 0.62, 0.51; the frequency of genotype AB was 0.06, 0.34, 0.34; the frequency of genotype BB was 0.01, 0.04, 0.15, respectively. The Small Tail Han ewes with genotype BB or AB had 0.64-0.76 or 0.44-0.54 more lambs than those with genotype AA, respectively. These results preliminarily showed that the prolactin receptor locus is either a major gene that influences the prolificacy in Small Tail Han sheep or is in close linkage with such a gene.     

Prolactin Receptor as a Candidate Gene for Prolificacy of Small Tail Han Sheep

DNA polymorphism of the ovine prolactin receptor gene (PRLR) was investigated and used to study its effect on litter size in sheep. By means of PRLR gene sequence homology between sheep and human, three primer pairs were designed for polymerase chain reaction (PCR) amplification within intron 1 and exon 10 of the PRLR gene in sheep. In these parts of the gene the single nucleotide polymorphisms were detected by PCR-single strand conformation polymorphism (SSCP) in 314 Small Tail Han ewes. These poly-morphisms were used to study the associations with litter size. The results indicated that there were three genotypes (AA, AB and BB) detected by three primer pairs. For three primer pairs the frequency of allele A was 0.96, 0.79, 0.68; and the frequency of allele B was 0.04, 0.21, 0.32, respectively. The frequency of genotype AA was 0.93, 0.62, 0.51; the frequency of genotype AB was 0.06, 0.34, 0.34; the frequency of genotype BB was 0.01, 0.04, 0.15, respectively. The Small Tail Han ewes with genotype BB or AB had 0.64–0.76 or 0.44–0.54 more lambs than those with genotype AA, respectively. These results preliminarily showed that the prolactin receptor locus is either a major gene that influences the prolificacy in Small Tail Han sheep or is in close linkage with such a gene.     

A/G Gln20Arg (exon 1) and G/A Val156Met (exon 5) polymorphisms of the human orosomucoid 1 gene in Mexico

The human orosomucoid 1 gene (ORM1) codes an alpha-1-acid glycoprotein that has been classified as an acute-phase reactive protein, and a major drug-binding serum component, as well as an immunomodulatory protein with genetic polymorphisms. Evaluation of ORM variation through isoelectric focusing and immunobloting has revealed a world-wide distribution of the ORM1 F and ORM1 S alleles. We evaluated and examined the genetic characteristics of two Mexican populations that have different anthropological and cultural antecedents, examining two ORM1 genotypes (exon 1 - A/G (Gln20Arg) and exon 5 G/A (Val156Met)) in 145 individuals, using nested polymerase chain reaction, sequencing, and restricted fragment length polymorphism. Mexican Mestizos had higher frequencies of the exon 1 A allele (P = 0.020) and AA genotype (P = 0.018) and lower frequency of the G allele (P = 0.020) when compared to Teenek Amerindians. When we examined exon 5 G/A (Val156Met) polymorphisms, we found significantly higher frequencies of the G allele (P = 0.0007) and the GG genotype (P = 0.0003) in the Mexican Mestizo population. The Teenek population had a significantly higher frequency of the A allele than has been reported for Chinese and African (P < 0.05) populations, and the G/A genotype was more frequently found in this Mexican population than in Chinese, African and European populations (P < 0.05).     

Single nucleotide polymorphisms of the prolactin receptor (PRLR) gene and its association with growth traits in chinese cattle

Genetic polymorphism of the prolactin receptor (PRLR) gene was detected by PCR-SSCP and DNA sequencing methods in 665 individuals from five Chinese cattle breeds. The results showed that at the P1 locus, three observed genotypes (AA, AB and BB), two linked SNPs (G1267A and T1268C), and one missense mutation (S18N) within a putative signal peptide were determined. The frequencies of haplotypes A and B in the five breeds were 0.596–0.802 and 0.198–0.404, respectively. Polymorphism of the PRLR gene was shown to be significantly associated with growth traits in the Nanyang breed. Individuals with genotype BB had greater hucklebone width, body weight and average daily gain than those with genotype AA at 6 months old (P < 0.01), as well as better body height, body length and heart girth when 6 months (P < 0.05). This study revealed for the first time that the PRLR gene is a promising candidate gene that affects growth traits in cattle.     

2010年深圳地区诺如病毒的基因分型

了解2010年深圳地区诺如病毒的基因型别及分子流行病学特点。 用诺如病毒特异性引物（GI-SKF/GI-SKR、COG2F/G2-SKR）,通过逆转录-聚合酶链反应（RT-PCR）方法进行诺如病毒核酸扩增,阳性产物回收纯化并测序,用Clustal W和MEGA4.0生物软件对诺如病毒序列进行序列比对和系统进化分析。 85份阳性标本中有79株诺如GⅡ型和6株诺如GⅠ型,其中55株为GⅡ/4(2006b)型,16株为GⅡ/4(2008variant)型,2株为GⅡ/1型,4株为GⅡ/5型,2株为GⅡ/11型,1株为GI/4型,2株为GI/5型,3株为GI/6型。 2010年深圳地区诺如病毒的主要型别是GI和GⅡ,并且以GⅡ/4型为主,流行优势株为GⅡ/4（2006b）。     

Molecular Characterization of Human Respiratory Syncytial Virus in the Philippines, 2012-2013

Human respiratory syncytial virus (HRSV) is a major cause of acute lower respiratory tract infections in infants and children worldwide. We performed molecular analysis of HRSV among infants and children with clinical diagnosis of severe pneumonia in four study sites in the Philippines, including Biliran, Leyte, Palawan, and Metro Manila from June 2012 to July 2013. Nasopharyngeal swabs were collected and screened for HRSV using real-time polymerase chain reaction (PCR). Positive samples were tested by conventional PCR and sequenced for the second hypervariable region (2^nd HVR) of the G gene. Among a total of 1,505 samples, 423 samples were positive for HRSV (28.1%), of which 305 (72.1%) and 118 (27.9%) were identified as HRSV-A and HRSV-B, respectively. Two genotypes of HRSV-A, NA1 and ON1, were identified during the study period. The novel ON1 genotype with a 72-nucleotide duplication in 2^nd HVR of the G gene increased rapidly and finally became the predominant genotype in 2013 with an evolutionary rate higher than the NA1 genotype. Moreover, in the ON1 genotype, we found positive selection at amino acid position 274 (p<0.05) and massive O- and N-glycosylation in the 2^nd HVR of the G gene. Among HRSV-B, BA9 was the predominant genotype circulating in the Philippines. However, two sporadic cases of GB2 genotype were found, which might share a common ancestor with other Asian strains. These findings suggest that HRSV is an important cause of severe acute respiratory infection among children in the Philippines and revealed the emergence and subsequent predominance of the ON1 genotype and the sporadic detection of the GB2 genotype. Both genotypes were detected for the first time in the Philippines.     

Genetic polymorphism in GST, NAT2, and MTRR and susceptibility to childhood acute leukemia

It is known that presence of xenobiotic-metabolizing gene polymorphisms in some cases correlates with hereditary predisposition to the oncological diseases. In the present work the frequencies of xenobiotic-metabolizing gene polymorphisms in 332 children with the diagnosis acute lymphoblastic leukemia (ALL), 71 children with the diagnosis acute myeloblastic leukemia (AML) and 490 healthy donors have been determined using allele-specific hybridization on the biochip. Statistically significant increase in the frequency of GSTT1 "null" genotype (OR = 1.9, p = 4.7E-5) and GSTT1/GSTM1 double "null" genotype (OR = 3.1, p = 2.5E-8) in children with acute leukemia relative to healthy donors group has been revealed. Also 1.8-fold increase in the frequency of NAT2 genotype 341T/T, 481C/C, 590G/G in children with acute leukemia relative to healthy donors group (p = 0.026) has been recognized. Analysis of gene-gene interactions has showed that in patients with acute leukemia genotype NAT2 341T/T, 481C/C, 590G/G in combination with GSTT1 "null" and/or GSTM1 "null" genotype is significantly more frequent than in population control. Besides the reduction of MTRR genotype 66G/G frequency in girls with acute leukemia relative to female healthy donors has been found (OR = 0.50, p = 0.0015). Analysis of gene-gene interactions has shown that the presence of GSTT1 "null" and/or GSTM1 "null" genotype in combination with MTRR genotype 66A/- may consider as risk factor of acute leukemia in girls. Thus, the studied polymorphic variants of genes GSTT1, GSTM1, NAT2 and MTRR can modulate the risk of childhood acute leukemia, residents of European part of Russia.