An integrative study of the temperature dependence of whole animal and muscle performance during jumping and swimming in the frog Rana temporaria

The aims of this study were: (1) to analyze individual variation in frog locomotor performance, (2) to compare the thermal sensitivity of jumping and swimming, and (3) to contrast whole animal versus muscle fiber performance at different temperatures. The jumping and swimming performance of Rana temporaria was analyzed at 5, 10, 15 and 20 °C. Muscle fiber bundles were isolated from lateral gastrocnemius and subjected to the length and activation patterns thought to occur in vivo. As temperature increased, locomotor performance in R. temporaria improved with a Q ₁₀ of 1.2 for both jump take-off velocity and mean swimming velocity. The slope of the relationship between performance and temperature (TE) was similar for both locomotor parameters and was described by the equation z-scores of locomotor performance = 0.127 × TE − 1.585. Although some frogs performed better than others relative performance was affected by locomotor type and temperature. Locomotor performance improved with temperature as the power required during take-off and the mean muscle power output increased with Q ₁₀ values of 1.7 and 1.6 respectively. The mean muscle power output during take-off was only 34% of the calculated requirements for the whole animal, suggesting the involvement of elastic strain energy storage mechanisms. Accepted: 2 September 1999     

Muscle activity in the leg is tuned in response to ground reaction forces.

During walking and running, the human body reacts to its external environment. One such response is to the impact forces that occur at heel strike. This study tested previous speculation that the levels of muscle activity in the lower extremities are adjusted in response to the loading rate of the impact forces. A pendulum apparatus was used to deliver repetitive impacts to the heels of 20 subjects. Impact forces were of similar magnitude to those experienced during running, but the loading rate was varied by 13% using different materials in the subjects' shoes. Myoelectric patterns were measured in the tibialis anterior, medial gastrocnemius, vastus medialis, and biceps femoris muscles. Wavelet analysis was used to resolve intensity of the myoelectric patterns into time and frequency space. Substantial and significant differences in the myoelectric activity occurred between the impact conditions for the 50 ms before and the 50 ms after impact, reaching 3 ms in timing, 16% in wavelet number, and 154% in the intensity of the muscle activity.     

Intracellular phosphorylation of benzyladenosine is related to apoptosis induction in tobacco BY-2 cells

Treatment of tobacco BY‐2 cells with micromolar concentration of benzyladenosine ([9R]BA) resulted in the loss of cell viability in a time‐ and concentration‐dependent manner. Cell death induced by [9R]BA exhibited typical apoptotic hallmarks including cell shrinkage, chromatin condensation and degradation of nuclear DNA to characteristic high molecular weight (HMW) as well as nucleosomal size fragments. Externally added [9R]BA was very rapidly and almost quantitatively phosphorylated within BY‐2 cells. Accumulation of [9R]BA‐monophosphate was accompanied by massive production of endogenous reactive oxygen species (ROS), intracellular ATP depletion, and these events were followed by the loss of cell viability. Inhibition of intracellular phosphorylation of [9R]BA by adenosin kinase inhibitor, 5′‐amino‐5′‐deoxyadenosine (AdAs), diminished ROS production, ATP depletion, and consequently prevented cells from death. Selective inhibition of ROS production without restoring ATP production, however, did not provide any protection to cells. In contrast, even enhanced phosphorylation of [9R]BA caused by adenosine that simultaneously revived ATP synthesis reduced the number of dying cells. This is the first evidence of a direct relationship between intracellular phosphorylation of [9R]BA and apoptosis induction in BY‐2 cells. ATP depletion but not ROS production is the key secondary event that determines the cellular decision between life and death.     

Vibrio cholerae laboratory infection of the adult house fly Musca domestica

The present study was designed to test the hypothesis that house flies may be capable of specifically harbouring ingested Vibrio cholerae in their digestive tracts. Flies were continuously fed green fluorescent protein (GFP)‐labelled, non‐O1/non‐O139 environmental strains of V. cholerae. Bacterial burdens were quantitatively measured using plate counts and localization was directly observed using confocal microscopy. Vibrio cholerae were present in the fly alimentary canal after just 4 h, and reached a plateau of ～10⁷ colony‐forming units (CFU)/fly after 5 days in those flies most tolerant of the pathogen. However, individual flies were resistant to the pathogen: one or more flies were found to carry < 180 V. cholerae CFU at each time‐point examined. In flies carrying V. cholerae, the pathogen was predominantly localized to the midgut rather than the rectal space or crop. The proportion of house flies carrying V. cholerae in the midgut was dose‐dependent: the continuous ingestion of a concentrated, freshly prepared dose of V. cholerae increased the likelihood that fluorescent cells would be observed. However, V. cholerae may be a transient inhabitant of the house fly. This work represents the first demonstration that V. cholerae can inhabit the house fly midgut, and provides a platform for future studies of host, pathogen and environmental mediators of the successful colonization of this disease vector.     

Biology and mode of action of pure antioestrogens

The properties of a series of 7 alpha-alkyl analogues of oestradiol are described. Studies of chemical structure and activity in the immature rat uterotrophic/antiuterotrophic assay revealed that molecules containing a terminal functional group (acid, alcohol, amine, amide) linked to the steroid by a decamethylene bridge possess both oestradiol agonist and antagonist activity. However, certain amides, exemplified by the compound ICI 164,384 [N-n-butyl-11-(3,17 beta-dihydroxyoestra-1,3,5(10)-trien-7 alpha-yl)-N-methylundecanamide], were devoid of oestrogenic activity but possessed potent antioestrogenic activity. Comparison of receptor binding and biological potency of steroid 7 alpha- and 7 beta-isomers showed that activity is confined largely to the 7 alpha-isomer. Comparison of the effects of tamoxifen and ICI 164,384 on progesterone receptor (PR) concentration in the rat uterus showed that, unlike tamoxifen, ICI 164,384 did not induce PR and blocked induction of PR by oestradiol. Chronic treatment of mature female rats with ICI 164,384 led to an ovariectomy-like regression of the uterus without affecting LH secretion or the rate of growth. ICI 164,384 was also an effective antitumour agent in rats bearing carcinogen-induced mammary tumours.     

SavvyCNV: Genome-wide CNV calling from off-target reads

Identifying copy number variants (CNVs) can provide diagnoses to patients and provide important biological insights into human health and disease. Current exome and targeted sequencing approaches cannot detect clinically and biologically-relevant CNVs outside their target area. We present SavvyCNV, a tool which uses off-target read data from exome and targeted sequencing data to call germline CNVs genome-wide. Up to 70% of sequencing reads from exome and targeted sequencing fall outside the targeted regions. We have developed a new tool, SavvyCNV, to exploit this ‘free data’ to call CNVs across the genome. We benchmarked SavvyCNV against five state-of-the-art CNV callers using truth sets generated from genome sequencing data and Multiplex Ligation-dependent Probe Amplification assays. SavvyCNV called CNVs with high precision and recall, outperforming the five other tools at calling CNVs genome-wide, using off-target or on-target reads from targeted panel and exome sequencing. We then applied SavvyCNV to clinical samples sequenced using a targeted panel and were able to call previously undetected clinically-relevant CNVs, highlighting the utility of this tool within the diagnostic setting. SavvyCNV outperforms existing tools for calling CNVs from off-target reads. It can call CNVs genome-wide from targeted panel and exome data, increasing the utility and diagnostic yield of these tests. SavvyCNV is freely available at https://github.com/rdemolgen/SavvySuite.     

A comparative study of the interaction of oestradiol and the steroidal pure antioestrogen, ICI 164,384, with the molybdate-stabilized oestrogen receptor

The kinetics of binding of oestradiol and the steroidal pure antioestrogen ICI 164,384 to the molybdate-stabilized oestrogen receptor, partially purified from pig and human uterine tissue, were determined. ICI 164,384 bound directly to the oestrogen receptor protein and the kinetic parameters of this interaction were, in general, similar to those for the binding of oestradiol, regardless of the source of the receptor protein. However, the rate of association of oestradiol, regardless of the source of the receptor protein. However, the rate of association of the antagonist with the receptor protein was slower when compared to that of oestradiol. Furthermore, the concentration of binding sites for the two ligands was of the same order. The binding of oestradiol resulted in a steroid-receptor complex which could be transformed in vitro, to a form with increased affinity for DNA-cellulose. However, the complex formed between ICI 164,384 and the receptor protein did not show increased affinity for DNA-cellulose when exposed to conditions that transformed agonist-receptor complexes. Therefore, the binding of ICI 164,384 to the oestrogen receptor protein results in a suppression of the transformation process. A similar suppression in vivo may account for the pure antagonist properties of ICI 164,384.