Hyperphenylalaninaemias in Estonia: Genotype-Phenotype Correlation and Comparative Overview of the Patient Cohort Before and After Nation-Wide Neonatal Screening.

The present study provides a retrospective overview of the cohort of phenylketonuria (PKU) patients in Estonia. Based on the available data, the patients clearly cluster into two distinct groups: the patients with late diagnosis and start of therapy (N = 46), who were born before 1993 when the national newborn screening programme was launched, and the screened babies (N = 48) getting their diagnoses at least in a couple of weeks after birth.Altogether 153 independent phenylalanine hydroxylase (PAH) alleles from 92 patients were analysed in the study, wherein 80% of them were carrying the p.Arg408Trp variation, making the relative frequency of this particular variation one of the highest known. Additionally, 15 other different variations in the PAH gene were identified, each with very low incidence, providing ground for phenotypic variability and potential response to BH4 therapy. Genealogical analysis revealed some "hotspots" of the origin of the p.Arg408Trp variation, with especially high density in South-East Estonia. According to our data, the incidence of PKU in Estonia is estimated as 1 in 6,700 newborns.

The actual role of oxygen deficit in the linkage of the water quality and benthic phosphorus release: Potential implications for lake restoration.

Human activities in watersheds have resulted in huge accumulations of phosphorus (P) in sediments that have subsequently hindered restoration efforts of lake water quality managers worldwide. Much controversy exists about the factors that control the release of P from sediments (internal P loading). One of the main debates concerns the role of oxygen deficit (anoxia) in the regulation of water quality. Our results based on a comprehensive set of lakes worldwide demonstrate that internal P loading (IPtot) plays a significant role in water quality regulation. Internal P loading due to anoxia (IPanox) contributes significantly to the IPtot. However, this contribution is insufficient to significantly increase the chlorophyll a (Chl a) concentration in stratifying lakes. In the lakes of the north temperate and boreal zone, this is because the IPanox reaches surface water layer in the end of the growing season. Observed water quality implications of IPtot are most likely caused by the sedimentary P that actually originates from the shallow areas. These findings suggest limitations for the use of aeration (improvement of the oxygen conditions in the hypolimnion) in lake water quality restoration. Moreover, lake ecosystem managers can benefit from our model that enables to predict anoxia triggered sedimentary P release from the combination of lake characteristics. The final decision on the use of aeration is indeed unique to each lake, and lake specific targets should be considered.

Prevalence of c.35delG and p.M34T mutations in the GJB2 gene in Estonia.

OBJECTIVE: The purpose of this study was to determine the prevalence of c.35delG and p.M34T mutations in the GJB2 gene among children with early onset hearing loss and within a general population of Estonia. METHODS: Using an arrayed primer extension assay, we screened 233 probands with early childhood onset hearing loss for 107 different mutations in the GJB2 gene. We then looked for the two most common mutations, c.35delG and p.M34T, in a population of 998 consecutively born Estonian neonates to determine the frequency of these mutations in the general population. RESULTS: In 115 (49%) of the patients with early onset hearing loss, we found a mutation in at least one allele of the GJB2 gene. Seventy-three (31%) were homozygous for the c.35delG mutation, seven (3%) were homozygous for the p.M34T mutation, and five (2%) had c35delG/p.M34T compound heterozygosity. Other six identified mutations in GJB2 gene occurred rarely. Among the 998 anonymous newborn samples, we detected 45 who were heterozygous for c.35delG, 2 individuals homozygous for c.35delG, and 58 who were heterozygous for p.M34T. Additionally, we detected two c.35delG/p.M34T compound heterozygotes. CONCLUSION: The most common GJB2 gene mutations in Estonian children with early onset hearing loss were c.35delG and p.M34T, with c.35delG accounting for 75% of GJB2 alleles. The carrier frequency for c.35delG and p.M34T in a general population of Estonia was 1 in 22 and 1 in 17, respectively, and was higher than in most other countries.

Predicting failure rate of PCR in large genomes.

We have developed statistical models for estimating the failure rate of polymerase chain reaction (PCR) primers using 236 primer sequence-related factors. The model involved 1314 primer pairs and is based on more than 80 000 PCR experiments. We found that the most important factor in determining PCR failure is the number of predicted primer-binding sites in the genomic DNA. We also compared different ways of defining primer-binding sites (fixed length word versus thermodynamic model; exact match versus matches including 1-2 mismatches). We found that the most efficient prediction of PCR failure rates can be achieved using a combination of four factors (number of primer-binding sites counted in different ways plus GC% of the primer) combined into single statistical model GM1. According to our estimations from experimental data, the GM1 model can reduce the average failure rate of PCR primers nearly 3-fold (from 17% to 6%). The GM1 model can easily be implemented in software to premask genome sequences for potentially failing PCR primers, thus improving large-scale PCR-primer design.

Influence of oocytes and spermatozoa on early embryonic development.

OBJECTIVE: To evaluate the effect of oocytes and spermatozoa on early embryonic development. DESIGN: Retrospective study. SETTING: Infertility Clinic, the Family Federation of Finland. PATIENT(S): Fifty-nine oocyte donation cycles with oocytes shared among 118 recipient couples. INTERVENTION(S): Culture of all fertilized oocytes. MAIN OUTCOME MEASURE(S): Standard sperm (concentration, progressive motility, and morphology according to Tygerberg strict criteria) and embryo (morphology and cleavage stage) characteristics. RESULT(S): A marked effect of the oocyte on both embryo morphology and blastomere cleavage rate was demonstrated. In addition, a significant sperm effect on blastomere cleavage rate was found. Sperm morphology as determined according to strict criteria rather than sperm count or progressive motility was positively associated with the blastomere cleavage rate. None of the measured sperm characteristics influenced embryo morphology. CONCLUSION(S): Embryo morphology, i.e., fragmentation and blastomere uniformity, are predominantly determined by oocyte quality, whereas both the oocyte and spermatozoa influence the blastomere cleavage rate.