Association of maternal serum Netrin-1 and Netrin-4 levels with placenta accreta spectrum.

OBJECTIVE: This study aimed to evaluate maternal serum Netrin-1 and Netrin-4 levels in pregnancies complicated with placenta accreta spectrum. PATIENTS AND METHODS: This cross-sectional study enrolled 49 pregnant women with the diagnosis of placenta accreta spectrum as the study group. Gestational age-matched 30 uncomplicated pregnant women with prior cesarean delivery and normal placentation were randomly selected as the control group. Maternal serum Netrin-1 and Netrin-4 levels were measured between weeks 34 and 36 of gestation. RESULTS: There was no significant difference between the groups in terms of demographic characteristics. Maternal serum Netrin-1 levels were significantly lower in placenta accreta spectrum cases compared with those in the control group (p=0.038). There was no significant difference between the groups in terms of maternal serum Netrin-4 levels (p>0.05). There was a significant negative correlation between maternal serum Netrin-1 levels and the number of prior cesarean deliveries (r=-0.313, p=0.005). CONCLUSIONS: The observed decrease in maternal serum Netrin-1 levels in placenta accreta spectrum cases associated with increased angiogenesis might be one of the factors involved in the pathophysiology of this disease.

A missense mutation in growth differentiation factor 9 (GDF9) is strongly associated with litter size in sheep.

BACKGROUND: A genome wide association study for litter size in Norwegian White Sheep (NWS) was conducted using the recently developed ovine 50K SNP chip from Illumina. After genotyping 378 progeny tested artificial insemination (AI) rams, a GWAS analysis was performed on estimated breeding values (EBVs) for litter size. RESULTS: A QTL-region was identified on sheep chromosome 5, close to the growth differentiation factor 9 (GDF9), which is known to be a strong candidate gene for increased ovulation rate/litter size. Sequencing of the GDF9 coding region in the most extreme sires (high and low BLUP values) revealed a single nucleotide polymorphism (c.1111G>A), responsible for a Val→Met substitution at position 371 (V371M). This polymorphism has previously been identified in Belclare and Cambridge sheep, but was not found to be associated with fertility. In our NWS-population the c.1111G>A SNP showed stronger association with litter size than any other single SNP on the Illumina 50K ovine SNP chip. Based on the estimated breeding values, daughters of AI rams homozygous for c.1111A will produce minimum 0.46 - 0.57 additional lambs compared to daughters of wild-type rams. CONCLUSION: We have identified a missense mutation in the bioactive part of the GDF9 protein that shows strong association with litter size in NWS. Based on the NWS breeding history and the marked increase in the c.1111A allele frequency in the AI ram population since 1983, we hypothesize that c.1111A allele originate from Finnish landrace imported to Norway around 1970. Because of the widespread use of Finnish landrace and the fact that the ewes homozygous for the c.1111A allele are reported to be fertile, we expect the commercial impact of this mutation to be high.

Correlation between broth microdilution and disk diffusion methods for antifungal susceptibility testing of caspofungin, voriconazole, amphotericin B, itraconazole and fluconazole against Candida glabrata.

Candida glabrata is one of the most frequent organisms isolated from superficial and invasive fungal infections, after Candida albicans. This organism also exhibits intrinsically low susceptibility to azole antifungals and treatment often fails. The microdilution method is not very practical for use in routine susceptibility testing in the clinical laboratory, thus necessitating the use of other methods. In this study, we compared the in vitro activity of five antifungal agents in three different groups (echinocandin, polyene and azole) against 50 C. glabrata isolates by broth microdilution and disk diffusion methods recommended by Clinical Laboratory Standards Institute CLSI M27-A3 and CLSI M44-A, respectively. All the isolates were susceptible to amphotericin B (100%) and 98% of the isolates were susceptible to caspofungin by the broth microdilution method. Within the azole group drugs, voriconazole was the most active followed by fluconazole and itraconazole in vitro. The highest rate of resistance was obtained against itraconazole with a high number of isolates defined as susceptible-dose dependent or resistant. Although the disk diffusion method is easy to use in clinical laboratories, it shows very poor agreement with the reference method for fluconazole and itraconazole against C. glabrata (8% and 14%, respectively).

A nonsense mutation in the beta-carotene oxygenase 2 (BCO2) gene is tightly associated with accumulation of carotenoids in adipose tissue in sheep (Ovis aries).

BACKGROUND: Sheep carcasses with yellow fat are sporadically observed at Norwegian slaughter houses. This phenomenon is known to be inherited as a recessive trait, and is caused by accumulation of carotenoids in adipose tissue. Two enzymes are known to be important in carotenoid degradation in mammals, and are therefore potential candidate genes for this trait. These are beta-carotene 15,15'-monooxygenase 1 (BCMO1) and the beta-carotene oxygenase 2 (BCO2). RESULTS: In the present study the coding region of the BCMO1 and the BCO2 gene were sequenced in yellow fat individuals and compared to the corresponding sequences from control animals with white fat. In the yellow fat individuals a nonsense mutation was found in BCO2 nucleotide position 196 (c.196C>T), introducing a stop codon in amino acid position 66. The full length protein consists of 575 amino acids. In spite of a very low frequency of this mutation in the Norwegian AI-ram population, 16 out of 18 yellow fat lambs were found to be homozygous for this mutation. CONCLUSION: In the present study a nonsense mutation (c.196C>T) in the beta-carotene oxygenase 2 (BCO2) gene is found to strongly associate with the yellow fat phenotype in sheep. The existence of individuals lacking this mutation, but still demonstrating yellow fat, suggests that additional mutations may cause a similar phenotype in this population. The results demonstrate a quantitatively important role for BCO2 in carotenoid degradation, which might indicate a broad enzyme specificity for carotenoids. Animals homozygous for the mutation are not reported to suffer from any negative health or development traits, pointing towards a minor role of BCO2 in vitamin A formation. Genotyping AI rams for c.196C>T can now be actively used in selection against the yellow fat trait.

Impact of two myostatin (MSTN) mutations on weight gain and lamb carcass classification in Norwegian White Sheep (Ovis aries).

BACKGROUND: Our aim was to estimate the effect of two myostatin (MSTN) mutations in Norwegian White Sheep, one of which is close to fixation in the Texel breed. METHODS: The impact of two known MSTN mutations was examined in a field experiment with Norwegian White Sheep. The joint effect of the two MSTN mutations on live weight gain and weaning weight was studied on 644 lambs. Carcass weight gain from birth to slaughter, carcass weight, carcass conformation and carcass fat classes were calculated in a subset of 508 lambs. All analyses were carried out with a univariate linear animal model. RESULTS: The most significant impact of both mutations was on conformation and fat classes. The largest difference between the genotype groups was between the wild type for both mutations and the homozygotes for the c.960delG mutation. Compared to the wild types, these mutants obtained a conformation score 5.1 classes higher and a fat score 3.0 classes lower, both on a 15-point scale. CONCLUSIONS: Both mutations reduced fatness and increased muscle mass, although the effect of the frameshift mutation (c.960delG) was more important as compared to the 3'-UTR mutation (c.2360G>A). Lambs homozygous for the c.960delG mutation grew more slowly than those with other MSTN genotypes, but had the least fat and the largest muscle mass. Only c.960delG showed dominance effects.

An insertion in the coding region of the myostatin (MSTN) gene affects carcass conformation and fatness in the Norwegian Spaelsau (Ovis aries).

BACKGROUND: A phenotype of increased muscle mass (IMM) and reduced fat, comparable to reported effects of deleterious mutations in the myostatin gene (MSTN), has been observed in the Norwegian Spaelsau breed. However, the genotyping of five AI rams producing descendants with this phenotype, failed to reveal any of the known functional MSTN mutations. FINDINGS: In the present study, the coding region of the MSTN gene was sequenced in a Spaelsau ram lamb with this particular phenotype. A one base-pair insertion mutation (c.120insA) producing a premature stop codon in amino acid position 49 was identified. The consequence of this mutation is that the bioactive carboxy-terminal end of the protein is not translated, and a completely non-functional myostatin protein is produced. Among the 98 available AI rams of this breed, all five individuals having descendants with this particular phenotype were found to be heterozygous for the c.120insA mutation. The probability that these five selected AI rams should be heterozygous carriers of the c.120insA mutation purely by chance was calculated to be 3.1 x 10-7. In total, 7 AI rams were found to be heterozygous carriers of c.120insA. The estimated breeding values (EBVs) for EUROP carcass conformation and fat class for these 7 individuals also points towards a strong phenotypic effect of this mutation. CONCLUSION: Based upon the completely deleterious effect this novel c.120insA mutation has on myostatin protein function, and the documented phenotypic effect of comparable MSTN mutations in Norwegian White Sheep and other species, we conclude that this mutation is the functional explanation underlying the IMM phenotype in Norwegian Spaelsau. The allele distribution among the 98 genotyped AI rams support this conclusion, and also suggests that c.120insA is the most common reason for this phenotype in the Norwegian Spaelsau breed.

The regulatory basis of melanogenic switching.

Melanin produced in follicular melanocytes is the major basis for pigmentation of hair and wool in mammals. Two major types of melanin may be synthesized, the black/brown eumelanin and the reddish/yellow pheomelanin. Based on available cell biological evidence and reasonable assumptions, a mathematical model is developed to improve our understanding of melanogenic switching, i.e. the switching between eumelanin and pheomelanin production depending on the extracellular signalling context. In 1993, Ito proposed that melanogenic switching is due to the covalent binding of the intermediate DOPAquinone to the enzyme glutathione reductase. We were only able to obtain a good fit to available experimental data on the relation between pheomelanin levels and the activity of the key enzyme tyrosinase by taking Ito's hypothesis into account. Thus, our results support Ito's hypothesis, and suggest that melanogenic switching may be due to a jump between two stable production pattern states when the tyrosinase activity varies between two bifurcation levels. This implies that small changes in the levels of external regulatory factors may cause an accentuated change in the proportion of the produced colour pigments and may explain the fact that mammalian coat patterns often exhibit sharply delimited patches of either black or reddish colour.

Reliability and validity of the Turkish version of the Hamilton Depression Rating Scale.

The aim of the study was to examine the reliability and validity of the Turkish version of the Hamilton Depression Rating Scale (HDRS). Ninety-four patients with major depression/depressive mood disorders and 40 healthy controls participated in the study. The severity of depression was assessed with the HDRS, Beck Depression Inventory (BDI), and Clinical Global Impression score (CGI). The test-retest reliability coefficient of the HDRS was based on a 5-day interval was.85, with a Cronbach alpha coefficient of.75 and a split-half reliability coefficient of.76. Interrater reliability coefficients based on the independent ratings of four assessors were between.87 and.98. The correlation between the HDRS and BDI scores was.48, and between the HDRS and CGI it was.56. Principal Components Analysis yielded six factors. The correlation (-.13) between the control and patient groups indicates that the HDRS assesses depression very well.