Viability of bovine demi- and quarter-embryos after transfer

The viability of bovine demi- and quarter-embryos was investigated. Early compacting morulae were nonsurgically flushed from superovulated donor cows and were bisected by two microneedles. One of the halves was then split further into two quarters. Each demi- and quarter-embryo was placed in an evacuated zona pellucida. One demi- or two quarter-embryos were transferred non-surgically into cow or heifer recipients. Viability was measured by ultrasound scanning of the fetuses on Days 35, 48 and 60 of pregnancy. The pregnancy rates at Day 60 were 46.2% (6 13 ) for heifers and 33.3% (4 12 ) for cows after the transfer of a single demi-embryo. The transfer of two quarter-embryos resulted in a pregnancy rate of 61.5% (8 13 ) for heifers and 8.3% (1 12 ) for cows. Seven (53.8%) and four (33.3%) live fetuses were found on Day 60 following the transfer of demi-embryos into heifers and cows, respectively. The transfer of quarter-embryos resulted in 10 fetuses (38.5%) in the heifer recipients and only one fetus (4.2%) in the cow recipients. The results of this study suggest that heifers are more suitable than cows as recipients for quarter-embryos.     

Oligochaeta of Lake Taimyr: a preliminary survey

Lake Taimyr in Siberia is northernmost among the world's large lakes: 73°40–75°20N, 99–106°E. The lake area is up to 4650 km² in summer, with a maximum depth of 26 m and a mean depth of only 2.8 m. The ice-free period lasts about three months. The water level sinks 5.5–6 m during winter, so that 85% of the bottom surface is frozen into ice for some time and subjected to negative temperatures, probably down to –20 °C. In artificially melted sediment samples, 75–92% of animals survived. The average summer biomass of zoobenthos is about 1 g m^–2 wet weight, a half of this being formed by Oligochaeta. Altogether 76 samples with 3742 oligochaete specimens collected by V. N. Grëze in 1943–1944 were studied. At least 14 taxa of Tubificidae, Lumbriculidae, and Enchytraeidae were found in the lake, and some more enchytraeids in an adjacent river. Many immature animals could not be identified to species. Naididae were completely lacking probably due to the absence of macrovegetation. The shallow freezing zone is inhabited mostly by Alexandrovia ringulata. The profundal fauna is dominated by Lamprodrilus isoporus, Stylodrilus sp., and Isochaetides sp.     

The Sex Specific Genetic Variation of Energetics in Bank Voles,Consequences of Introgression?

Interaction between mitochondrial and nuclear genomes is expected to affect energetic phenotypes of traits linked to mitochondrial physiology, further influencing the fitness. A rodent, the bank vole (Myodes glareolus), has a population structure completely or partially introgressed with mitochondria from its relative, the red vole (M. r utilus). Females that carried either bank vole mitochondria or mitochondria from the introgressed species were repeatedly mated with males of both mtDNA types. We found that in males, but not in females, morpho-physiological phenotypes are affected by sire type, causing decreases in body mass (BM) and basal metabolic rate (BMR; including BM corrected, rBMR) in individuals sired by fathers carrying introgressed mitochondria. Higher effect sizes for the proportion of additive genetic variation (and 5.6, 1.9 and 3.6 times higher narrow sense heritability for BM, BMR and rBMR, respectively), and lower for proportion of environmental variation were detected in progeny of non-introgressed males. Our data indicate that co-adapted and possibly co-introgressed nuclear genes related to energetic physiology have an important role in adaptation to the northern conditions in bank voles, and that sex linked nuclear genes are a potential source for variation in basal metabolic rate.     

Using the nonlinear control of anaesthesia-induced hypersensitivity of EEG at burst suppression level to test the effects of radiofrequency radiation on brain function

Background  

In this study, investigating the effects of mobile phone radiation on test animals, eleven pigs were anaesthetised to the level where burst-suppression pattern appears in the electroencephalogram (EEG). At this level of anaesthesia both human subjects and animals show high sensitivity to external stimuli which produce EEG bursts during suppression. The burst-suppression phenomenon represents a nonlinear control system, where low-amplitude EEG abruptly switches to very high amplitude bursts. This switching can be triggered by very minor stimuli and the phenomenon has been described as hypersensitivity. To test if also radio frequency (RF) stimulation can trigger this nonlinear control, the animals were exposed to pulse modulated signal of a GSM mobile phone at 890 MHz. In the first phase of the experiment electromagnetic field (EMF) stimulation was randomly switched on and off and the relation between EEG bursts and EMF stimulation onsets and endpoints were studied. In the second phase a continuous RF stimulation at 31 W/kg was applied for 10 minutes. The ECG, the EEG, and the subcutaneous temperature were recorded.     

Real-Time Correlates of Phonological Quantity Reveal Unity of Tonal and Non-Tonal Languages

Discrete phonological phenomena form our conscious experience of language: continuous changes in pitch appear as distinct tones to the speakers of tone languages, whereas the speakers of quantity languages experience duration categorically. The categorical nature of our linguistic experience is directly reflected in the traditionally clear-cut linguistic classification of languages into tonal or non-tonal. However, some evidence suggests that duration and pitch are fundamentally interconnected and co-vary in signaling word meaning in non-tonal languages as well. We show that pitch information affects real-time language processing in a (non-tonal) quantity language. The results suggest that there is no unidirectional causal link from a genetically-based perceptual sensitivity towards pitch information to the appearance of a tone language. They further suggest that the contrastive categories tone and quantity may be based on simultaneously co-varying properties of the speech signal and the processing system, even though the conscious experience of the speakers may highlight only one discrete variable at a time.     

Probing pH-dependent functional elements in proteins: modification of carboxylic acid pairs in Trichoderma reesei cellobiohydrolase Cel6A

Two carboxylic acid side chains can, depending on their geometry and environment, share a proton in a hydrogen bond and form a carboxyl-carboxylate pair. In the Trichoderma reesei cellobiohydrolase Cel6A structure, five carboxyl-carboxylate pairs are observed. One of these pairs (D175-D221) is involved in catalysis, and three other pairs are found in, or close to the two surface loops covering the active site tunnel of the catalytic domain. To stabilize Cel6A at alkaline pH values, where deprotonation of the carboxylic acids leads to repulsion of their side chains, we designed two mutant enzymes. In the first mutant, one carboxyl-carboxylate pair (E107-E399) was replaced by a corresponding amide-carboxylate pair (Q107-E399), and in the second mutant, all three carboxyl-carboxylate pairs (E107-E399, D170-E184, and D366-D419) were mutated in a similar manner. The unfolding studies using both intrinsic tryptophan fluorescence and far-ultraviolet circular dichroism spectroscopy at different pH values demonstrate that the unfolding temperature (T(m)) of both mutants has changed, resulting in destabilization of the mutant enzymes at acidic pH and stabilization at alkaline pH. The effect of stabilization seems additive, as a Cel6A triple mutant is the most stable enzyme variant. This increased stability is also reflected in the 2- or 4-fold increased half-life of the two mutants at alkaline pH, while the catalytic rate on cellotetraose (at t = 0) has not changed. Increased operational stability at alkaline pH was also observed on insoluble cellulosic substrates. Local conformational changes are suggested to take place in the active site loops of Cel6A wild-type enzyme at elevated pHs (pH 7), affecting to the end-product spectrum on insoluble cellulose. The triple mutant does not show such pH-dependent behavior. Overall, our results demonstrate that carboxyl-carboxylate pair engineering is a useful tool to alter pH-dependent protein behavior.     

Nerve growth factor in serum of children with systemic lupus erythematosus is correlated with disease activity

Nerve growth factor (NGF) is a neurotrophic factor, which is expressed both in the nervous system and in peripheral organs. NGF is also present in mast cells, and in B- and T-lymphocytes, and may play a role in the immune cell development and differentiation. Various cytokines have been shown to affect NGF expression, and NGF is elevated in inflammation and in some autoimmune diseases. Here we have studied NGF concentrations in serum of pediatric patients with systemic lupus erythematosus (SLE) using a two-site enzyme-linked immunosorbent assay (ELISA). We have further correlated the levels of NGF to the inflammatory state of the disease. The mean value of serum NGF in SLE patients was significantly increased compared with controls (3346 vs 627pg/ml). There was a correlation between the activity of SLE and the levels of NGF. The results show that NGF is elevated in childhood SLE and that the levels are correlated with disease activity. The present results suggest that NGF may play a role in the pathogenesis of SLE and may have a prognostic value in evaluating the course of the disease and in outlining the medication.     

Indicative profundal oligochaete assemblages in selected small Swedish lakes

Fifty-seven small- to middle-size lakes of Sweden (size-range from <0.1 to about 200 km²) representing different regions and lake-types were selected for the purpose of demonstrating the indicative value of profundal oligochaete communities also of relatively small lakes. It is well known since previously that profundal oligochaete assemblages of large lakes in Europe may be particularly indicative of the current trophic situation, but little has been known about smaller lakes in this respect. Characterizations based on oligochaete community structure in these lakes could be summarized in eight different groups: Characteristic oligochaete communities in oligotrophic lakes of relatively small size are Spirosperma ferox, Stylodrilus heringianus and Tubifex tubifex. Oligochaete species richness is low (1–3 species) and so is species abundance (<300 specimens m^–2). Very eutrophic lakes of the same size-range are largely dominated by the tubificids Limnodrilus hoffmeisteri and/or Potamothrix hammoniensis. Substitute or complementary species may be T. tubifex. Species richness is usually low (1–3 species), but abundance is high to very high (>5000 specimens m^–2). Mesotrophic to moderately eutrophic lakes of the same size categories contain more species than the above extremes, most of them being fairly tolerant to oxygene deficiency such as Tubifex ignotus, a number of species of the genera Aulodrilus and Limnodrilus and a variety of so-called Ponto-Caspian Potamothrix species, most of them having recently invaded south-eastern Sweden. Species richness is moderate to high (4–8 species), while abundance may vary considerably (range from about 300 to about 5000 specimens m^–2). In this respect, small- to middle-size lakes do not differ much from larger lakes. For the trophic characterization of lake water the mean concentrations of total-phosphorus (in g l^–1) is generally regarded to be the most appropriate measure. The match between characterizations based upon oligochaete community structure and total-phosphorus concentrations is generally very good for large lakes. The present study shows that this applies fairly well also to the majority of the small- to middle-size lakes selected for this study. This has not been shown before. Obvious deviations from this rule could be caused by a scarcity of reliable chemical data, or considerable seasonal shifts in the chemical composition ot the water, which in its turn may be caused by short water residence time. Littoral elements in the fauna, which are generally less indicative, or effects of local inflows may also be more frequent here than in lakes of larger size.     

Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport.

The yeast SEC1 gene encodes a hydrophilic protein that functions at the terminal stage in secretion. We have cloned two yeast genes, SSO1 and SSO2, which in high copy number can suppress sec1 mutations and also mutations in several other late acting SEC genes, such as SEC3, SEC5, SEC9 and SEC15. SSO1 and SSO2 encode small proteins with N-terminal hydrophilic domains and C-terminal hydrophobic tails. The two proteins are 72% identical in sequence and together perform an essential function late in secretion. Sso1p and Sso2p show significant sequence similarity to six other proteins. Two of these, Sed5p and Pep12p, are yeast proteins that function in transport from ER to Golgi and from Golgi to the vacuole, respectively. Also related to Sso1p and Sso2p are three mammalian proteins: epimorphin, syntaxin A/HPC-1 and syntaxin B. A nematode cDNA product also belongs to the new protein family. The new protein family is thus present in a wide variety of eukaryotic cells, where its members function at different stages in vesicular transport.     

Detection of microinjected genes in bovine preimplantation embryos with combined DNA digestion and polymerase chain reaction

We have developed a simple digestion-polymerase chain reaction (PCR) assay for a simultaneous transgene detection and sexing of pronucleus-injected bovine preimplantation embryos. Bovine embryos were microinjected with dam-methylated gene construct and cultured in vitro for 6–7 days after the injections. The developed blastocysts and compact morulae were bisected and the embryonic biopsies representing mainly trophoblasts were subjected to the digestion-PCR, while the biopsied embryos remained in culture. Embryonic DNA was released with proteinase K and the samples were digested with a Dpnl-Bal31 mixture before the PCR amplification of the transgene, bovine αS1-casein, and bovine Y-chromosome fragments in the same reaction. The whole assay from biopsy to electrophoresis took less than 6 hr. The digestion removed up to 50 fg of dam-methylated transgene copies (unintegrated or contaminants) and also a few hundred copies of contaminating PCR products from the embryonic samples. The digestion-PCR assay eliminated all transgene contaminations from noninjected blastocysts, which were exposed to the microinjection DNA during the stay in injection chambers, and reduced the amount of transgene-positive embryos among pronucleus-injected blastocysts as compared with unmodified PCR. Analysis of 486 microinjected bovine embryo biopsies in 13 separate experiments revealed that we were able to sex 398 (82%) of the biopsies and 77 (19%) of the biopsies were scored as transgene positive and 57 (14%) as transgene questionable. Upon reanalysis of 41 of the biopsied embryos, 38 (93%) of the embryos were observed to be transgene negative and 2 questionable in both assays and uneven distribution of transgene copies was observed in one embryo. The results from sexing were in accordance with biopsies and remaining embryos in 38 (93%) of the embryos. © 1996 Wiley-Liss, Inc.