Epidermal Growth Factor Gene (EGF) Polymorphism and Risk of Melanocytic Neoplasiay

A common single nucleotide polymorphism (SNP) in the 5' untranslated region (5'UTR) of the epidermal growth factor (EGF) gene modulates the level of transcription of this gene and hence is associated with serum levels of EGF. This variant may be associated with melanoma risk, but conflicting findings have been reported. An Australian melanoma case-control sample was typed for the EGF+61A>G transversion (rs4444903). The sample comprised 753 melanoma cases from 738 families stratified by family history of melanoma and 2387 controls from 645 unselected twin families. Ancestry of the cases and controls was recorded, and the twins had undergone skin examination to assess total body nevus count, degree of freckling and pigmentation phenotype. SNP genotyping was carried out via primer extension followed by matrix-assisted laser desorption time of flight (MALDI-TOF) mass spectroscopy. The EGIF+61 SNP was not found to be significantly associated with melanoma status or with development of nevi or freckles. Among melanoma cases, however, G homozygotes had thicker tumors (p=0.05), in keeping with two previous studies. The EGF polymorphism does not appear to predispose to melanoma or nevus development, but its significant association with tumor thickness implies that it may be a useful marker of prognosis.

Wetting and the physiological responses of grain-fed cattle in a heated environment

A controlled crossover experimental design was used to determine the effect of altered water sprinkling duration on heifers subjected to heat stress conditions. Heifers were subjected to 3 days of thermoneutral conditions followed by 3 days of hot conditions accompanied by water sprinkling between 1300 and 1500 h (HOT1-3). Then on the following 2 days (HOT4-5), environmental conditions remained similar, but 3 heifers were sprinkled between 1200 and 1600 h ( WET) and 3 were not sprinkled (NONWET). This was followed by a 1-day period (HOT6) in which environmental conditions and sprinkling regimen were similar to HOT1-3. Rectal temperature (RT) was collected hourly, and respiration rate (RR) was monitored every 2 h on HOT Days 2, 4, 5, and 6. Dry matter intake and rate of eating were also determined. Sprinkling reduced RR and RT (P < 0.01) of all heifers during HOT1-3. During HOT4-5, WET heifers had lower (P < 0.05) RT than NONWET from 1300 to 700 h and lower RR from 1400 to 2000 h. Dry matter intake of NONWET heifers was reduced by 30.6% (P < 0.05) during HOT4-5 and by 51.2% on HOT6. On HOT4-5 the dry matter intakes of WET heifers were similar to intakes under thermoneutral conditions. During HOT6, RT was again reduced following sprinkling in all heifers. Comparison of RT and RR of NONWET and WET heifers on HOT1-3 v. HOT6 revealed that under similar environmental conditions, NONWET heifers had increased RT, partially due to carry-over from HOT4-5. However, NONWET heifers had 40% lower feed intake but tended to have lower RR on HOT6 v. HOT1-3. Only RR of WET heifers was greater on HOT6, possibly a result of switching from a 4-h back to a 2-h sprinkling period, while maintaining a 62% greater intake (5.80 v. 3.58 kg/day) than NONWET heifers during this time. Results suggest that inconsistent cooling regimens may increase the susceptibility of cattle to heat stress and elicit different physiological and metabolic responses.

Novel variants in growth differentiation factor 9 in mothers of dizygotic twins

Context: Genes from the ovarian bone morphogenetic signaling pathway (GDF9 and BMP15) are critical for normal human fertility. We previously identified a deletion mutation in GDF9 in sisters with spontaneous dizygotic (DZ) twins, but the prevalence of rare GDF9 variants in twinning families is unknown. Objective: The objective was to evaluate the frequency of rare variants in GDF9 in families with a history of DZ twinning. Design and Subjects: We recruited 3450 individuals from 915 DZ twinning families (1693 mothers of twins) and 1512 controls of Caucasian origin. One mother of DZ twins was selected from 279 of the 915 families, and a DNA sample was screened for rare variants in GDF9 using denaturant HPLC. Variants were confirmed by DNA sequencing and genotyped in the entire sample by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. Results: We found two novel insertion/deletions (c.392-393insT, c.1268-1269delAA) and four missense alterations in the GDF9 sequence in mothers of twins. Two of the missense variants (c.307C > T, p.Pro103Ser and c.362C > T, p.Thr121Leu) were located in the proregion of GDF9 and two (c.1121C > T, p.Pro374Leu and c.1360C > T, p.Arg454Cys) in the mature protein region. For each variant, the frequencies were higher in cases compared with controls. The proportion of mothers of DZ twins carrying any variant (4.12%) was significantly higher (P < 0.0001) than the proportion of carriers in controls (2.29%). Conclusion: We describe new variants in the GDF9 gene that are significantly more common in mothers of DZ twins than controls, suggesting that rare GDF9 variants contribute to the likelihood of DZ twinning.

Growth of crystals in optical tweezers

We report here on the use of optical tweezers in the growth and manipulation of protein and inorganic crystals. Sodium chloride and hen egg-white lysozyme crystals were grown in a batch process, and then seeds from the solution were introduced into the optical tweezers. The regular and controllable shape and the known optical birefringence in these structures allowed a detailed study of the orientation effects in the beam due to both polarization and gradient forces. Additionally, we determined that the laser tweezers could be used to suspend a crystal for three-dimensional growth under varying conditions. Studies included increasing the protein concentration, thermal cycling, and a diffusion-induced increase in precipitant concentration. Preliminary studies on the use of the tweezers to create a localized seed for growth from polyethylene oxide solutions are also reported.