Gene targeting and somatic cell genetics--a rebirth or a coming of age?

A number of recent advances have significantly facilitated gene targeting in somatic cells. Gene targeting can now be performed with the same ease and efficiency in somatic cells as in murine embryonic stem cells. Rigorous genetic analyses can therefore be applied for the first time to the large number of excellent human cell culture systems. These tools will be important in areas where rodent models do not adequately represent human biology.     

Does transpiration control stomatal responses to water vapour pressure deficit?

Three types of observations were used to test the hypothesis that the response of stomatal conductance to a change in vapour pressure deficit is controlled by whole-leaf transpiration rate or by feedback from leaf water potential. Varying the leaf water potential of a measured leaf by controlling the transpiration rate of other leaves on the plant did not affect the response of stomatal conductance to vapour pressure deficit in Glycine max. In three species, stomatal sensitivity to vapour pressure deficit was eliminated when measurements were made at near-zero carbon dioxide concentrations, despite the much higher transpiration rates of leaves at low carbon dioxide. In Abutilon theophrasti, increasing vapour pressure deficit sometimes resulted in both decreased stomatal conductance and a lower transpiration rate even though the response of assimilation rate to the calculated substomatal carbon dioxide concentration indicated that there was no ‘patchy’ stomatal closure at high vapour pressure deficit in this case. These results are not consistent with stomatal closure at high vapour pressure deficit caused by increased whole-leaf transpiration rate or by lower leaf water potential. The lack of response of conductance to vapour pressure deficit in carbon dioxide-free air suggests that abscisic acid may mediate the response.     

Limitation of stomatal conductance by hydraulic traits: sensing or preventing xylem cavitation?

We tested the hypothesis that hydraulic conductance per unit leaf surface area of plant shoots (K_SL) determines the maximum diurnal stomatal conductance (g_L) that can be reached by plants growing in the field. A second hypothesis was tested that some xylem cavitation cannot be avoided by transpiring plants and might act as a signal for regulating g_L. Eleven woody species were studied, differing from each other with respect to taxonomy, wood anatomy and leaf habit. Maximum diurnal g_L, transpiration rate (E_L), pre-dawn and minimum diurnal leaf water potential (O_pd and O_min, respectively) were measured in the field. The critical O level at which stem cavitation was triggered (O_cav) was measured on detached branches, using the acoustic method. A high-pressure flow meter was used to measure maximum K_SL of 1-year-old shoots. Both g_L and E_L were positively related to K_SL. The whole-plant hydraulic conductance per unit leaf area (K_WL) of all the species studied, calculated as the ratio of E_L to (O (=O_pd-O_min) was closely related to K_SL. In every case, O_min (ranging between -0.85 and -1.35 MPa in the different species) dropped to the O_cav range or was <O_cav (ranging between -0.71 and -1.23 MPa), thus suggesting that some cavitation-induced embolism could not be avoided. The possibility is discussed that some cavitation-induced reduction in K_SL is the signal for stomatal closure preventing runaway embolism. The lack of correlation of g_L to O_cav is discussed in terms of the inconsistency of O_cav as an indicator of the vulnerability of plants to cavitation. No differences in hydraulic traits were observed between evergreen and deciduous species.     

Integrated control of vector-borne diseases of livestock--pyrethroids: panacea or poison?

Tick- and tsetse-borne diseases cost Africa approximately US$4-5 billion per year in livestock production-associated losses. The use of pyrethroid-treated cattle to control ticks and tsetse promises to be an increasingly important tool to counter this loss. However, uncontrolled use of this technology might lead to environmental damage, acaricide resistance in tick populations and a possible exacerbation of tick-borne diseases. Recent research to identify, quantify and to develop strategies to avoid these effects are highlighted.     

Headcase gene--proliferation or hormonal control?

A new insertion allele of the hdc gene was isolated and described. The nucleotide sequence of the coding region had no detectable homology with genomic DNA of any other Drosophila species, except for D. mauritiana. Gene expression was found both in adult testes and ovaries and at embryonic and larval stages. This expression pattern exhibits a strong similarity to that of cell-cycle genes. In contrast to cycE (a typical cell-cycle gene), which leads to expression termination after in vivo culturing of the wing disk, hdc did not arrest expression. It is concluded that hdc is a species-specific differentiation gene, whose regulatory activity in the development of an organism differs from that of proliferation genes.     

Dynamics of bacterioplankton in oligotrophic and eutrophic aquatic environments: bottom-up or top-down control?

Measurements of bacterial biomass, production and mortality have been carried out in a large range of aquatic environments, including eutrophic and oligotrophic ones. The general trends of variations of bacterial biomass, size, specific growth rate and mortality rate in all these environments are examined. The overall flux of bacterial production is taken as an index of the flux of organic matter available to bacteria, thus characterizing the richness of the environment. Bacterial biomass is roughly proportional to richness, while mean cell size increases with it. The turnover rate of biomass, as revealed either by growth or by mortality rates, appears to be fairly independent of richness.These observations are compatible with a simple resource-limited (bottom-up controlled) model of the dynamics of bacterioplankton. On the other hand, they are in contradiction with the predictions of a predator-controlled (top-down controlled) model.     

Does microbiological testing of foods and the food environment have a role in the control of foodborne disease in England and Wales?

This review looks at the contribution of microbiological sampling to the safety of retail foods in England and Wales. It compares sampling methods available and assesses the value of testing as part of outbreaks of foodborne disease, as part of routine management by local authorities, as part of work done or commissioned by the food industry, and as part of research. It confirms that microbiological testing has a role during outbreaks as it makes a significant contribution to help identify foods and other areas of greatest risk for future study. The review suggests that routine testing by local authorities is often of limited use and could be improved by more targeted surveillance. Testing could be better used to validate primary control methods, such as Hazard Analysis and Critical Control Point (HACCP) system. Any public health benefit from testing in the food industry is often restricted by client confidentiality. Microbial research on foods is important as it can lead to significant improvements in safety. Current microbiological methods are slow and, in future, rapid molecular methods may make an even bigger contribution to the control of foodborne disease.     

Clustering of insulin resistance, total and central abdominal fat: same genes or same environment?

Obesity, insulin resistance and disturbed glucose metabolism cluster within the Insulin Resistance Syndrome (IRS). Whether this reflects shared genetic or environmental factors detectable in 'normal' populations (not selected for IRS features) is unknown. This study estimated (i) genetic influences on IRS traits and (ii) shared and specific genetic and environmental factors on the relationships between these traits in healthy female twins. Fasting insulin, glucose, total and central fat were measured in 59 monozygotic (MZ) and 51 dizygotic (DZ) female twin pairs aged (+/- SD) 52 +/- 13 years. Body fat was measured by dual-energy X-ray absorptiometry, insulin resistance and secretion by a modified homeostasis model assessment. Using intraclass correlation coefficients and univariate model-fitting analyses, genetic influences were found in total fat, central fat, insulin resistance, fasting glucose and insulin secretion, with genetic factors explaining 64, 57, 59, 75 and 68% of their variance, respectively, using the latter technique. In matched analysis intra-pair differences in total and central fat related to intra-pair differences in insulin resistance (r2 = 0.19, P < 0.001). Multivariate model-fitting showed a close genetic relationship between total and central fat (r = 0.88). The genetic correlation between IR and central fat (0.41) was significantly greater than that for total fat (0.24), suggesting that central fat is not only a predictor of, but shares considerable genetic influence with, insulin resistance. In Cholesky analysis, these genetic influences were separate from those shared between central and total fat. In conclusion, both shared and specific genetic factors regulate components of the IRS in healthy females. However, there were discrete genetic influences on beta-cell insulin secretion, not shared with other IRS components, suggesting that a separate genetic propensity exists for Type 2 diabetes. These findings suggest we may understand the genetic and environmental influences on IRS from the study of the normal population.     

Mitotic control of dTTP pool: a necessity or coincidence?

The fidelity of DNA replication in eukaryotic cells requires a balanced dNTP supply in the S phase. During the cell cycle progression, the production of dTTP is highly regulated to coordinate with DNA replication. Intracellular thymidine is salvaged to dTTP by cytosolic thymidine kinase (TK1) and thymidylate kinase (TMPK), both of which expression increase in the G1/S transition and diminish in the mitotic phase via proteolytic destruction. Anaphase promoting complex/cyclosome (APC/C)-mediated ubiquitination targets TK1 and TMPK to undergo proteasomal degradation in mitosis, by which dTTP pool is minimized in the early G1 phase of the next cell cycle. In this review, we will focus on regulation of TK1 in the post-S phase and the importance of mitotic proteolysis in controlling dNTP balance, replication stress and genomic stability. Finally, we discuss how thymidine pool and oligomeric forms of TK1 can affect mitotic control of dTTP. This article is for the special issue of IMB 20^th anniversary.     

Femoral neck fragility: genes or environment?

As with other diseases that show exponentially increasing rates, hip fracture probably requires multiple prior events to become manifest. It is common ground that there are genetic, environmental, lifestyle and perhaps dietary determinants of risk of osteoporotic fracture as well as interactions between them. The key to secondary prevention is the understanding of how these components can be integrated into an effective assessment of the major risks of hip fracture after a previous fracture has occurred. The key to primary prevention is to understand both the pathological and physiological basis of hip fragility. It is not unreasonable to suppose that in a western lifestyle our limited and stereotypic patterns of locomotion from middle age onwards may offer considerably less protection than, for example, the more physically demanding activity of subsistence farming.     

Gene–gene and gene–environment interactions in ulcerative colitis

Genome-wide association studies (GWAS) have identified at least 133 ulcerative colitis (UC) associated loci. The role of genetic factors in clinical practice is not clearly defined. The relevance of genetic variants to disease pathogenesis is still uncertain because of not characterized gene–gene and gene–environment interactions. We examined the predictive value of combining the 133 UC risk loci with genetic interactions in an ongoing inflammatory bowel disease (IBD) GWAS. The Wellcome Trust Case–Control Consortium (WTCCC) IBD GWAS was used as a replication cohort. We applied logic regression (LR), a novel adaptive regression methodology, to search for high-order interactions. Exploratory genotype correlations with UC sub-phenotypes [extent of disease, need of surgery, age of onset, extra-intestinal manifestations and primary sclerosing cholangitis (PSC)] were conducted. The combination of 133 UC loci yielded good UC risk predictability [area under the curve (AUC) of 0.86]. A higher cumulative allele score predicted higher UC risk. Through LR, several lines of evidence for genetic interactions were identified and successfully replicated in the WTCCC cohort. The genetic interactions combined with the gene-smoking interaction significantly improved predictability in the model (AUC, from 0.86 to 0.89, P = 3.26E?05). Explained UC variance increased from 37 to 42 % after adding the interaction terms. A within case analysis found suggested genetic association with PSC. Our study demonstrates that the LR methodology allows the identification and replication of high-order genetic interactions in UC GWAS datasets. UC risk can be predicted by a 133 loci and improved by adding gene–gene and gene–environment interactions.     

"Lucy's" body height and relative leg length: human- or ape-like?

The body height of Australopithecus afarensis A.L. 288-1 ("Lucy") has recently been estimated and calculated as between 1 m to 1.06 m; other estimates give ca. 1.20 m. In addition, it is often stated that her relative leg length was shorter than that of modern humans. Using relative leg-, femur- and tibia length it is shown that both statements together can not be true; either her body height must at least have been around 1.06 to 1.10 m to give "Lucy" human-like leg proportions, or, to achieve a shorter, more ape-like leg ratio, a body height of ca. 1.20 m must be assumed.