Reduction of β-thiopyruvic acid by lactate dehydrogenase: a kinetic study

Summary In this paper a steady-state kinetic study on the system lactate dehydrogenase--thiopyruvate--thiolactate is presented and the possible mechanistic and physiological implications are discussed.At pH 7.4 the equilibrium between -thiopyruvate and -thiolactate, in the presence of NADH and lactate dehydrogenase is largerly shifted towards the formation of -thiolactate as in the case of pyruvate and lactate. This can be relevant in connection with the mixed disulfide between cysteine and -thiolactate that is observed to be present in the mammalian body fluids.The catalytic mechanism is of the bi-bi compulsory order type, and rapid equilibrium conditions for the binding of the first substrate (NADH) are shown to apply. A complex inhibition pattern of inhibitions by both substrates, however, prevents simple suggestions about the nature of the dead-end species involved.Abbreviations TPA -thiopyruvic acid - TLA -thiolactic acid - E enzyme This work is gratefully dedicated to Prof. Alessandro Rossi Fanelli on the occasion of his 75th birthday.     

Detection of a lactate threshold during incremental exercise?

An important question in the study of the exercise response is the real or imaginary nature of the anaerobic threshold, and mathematical modeling techniques have been invoked to assist in resolving this issue. Two opposing views with competing data models recently published in this journal are criticized. One view suggests a segmented model with a discontinuous first derivative at the threshold. The other suggests a continuous model over the whole work load range, implying the anaerobic threshold to be imaginary. However, neither group of authors has devoted proper rigorous attention to the models they use. Had this been done, some of the divergence of opinion may have been avoided. Ideal data from an alternate segmented model that has a continuous first derivative at the threshold are considered for comparative purposes. This suggests that the log-log transformation method may well lead to improved detection of a threshold when one exists, although the estimates of the threshold value obtained are unreliable. Modeling methodology is a useful approach to the resolution of scientific issues, but there exist fundamental implications and alternatives that must be fully recognized.     

Evidence for a stromal-epithelial “lactate shuttle” in human tumors

Recently, we proposed a new mechanism for understanding the Warburg effect in cancer metabolism. In this new paradigm, cancer-associated fibroblasts undergo aerobic glycolysis, and extrude lactate to “feed” adjacent cancer cells, which then drives mitochondrial biogenesis and oxidative mitochondrial metabolism in cancer cells. Thus, there is vectorial transport of energy-rich substrates from the fibroblastic tumor stroma to anabolic cancer cells. A prediction of this hypothesis is that cancer-associated fibroblasts should express MCT4, a mono-carboxylate transporter that has been implicated in lactate efflux from glycolytic muscle fibers and astrocytes in the brain. To address this issue, we co-cultured MCF7 breast cancer cells with normal fibroblasts. Interestingly, our results directly show that breast cancer cells specifically induce the expression of MCT4 in cancer-associated fibroblasts; MCF7 cells alone and fibroblasts alone, both failed to express MCT4. We also show that the expression of MCT4 in cancer-associated fibroblasts is due to oxidative stress, and can be prevented by pre-treatment with the anti-oxidant N-acetyl-cysteine. In contrast to our results with MCT4, we see that MCT1, a transporter involved in lactate uptake, is specifically upregulated in MCF7 breast cancer cells when co-cultured with fibroblasts. Virtually identical results were also obtained with primary human breast cancer samples. In human breast cancers, MCT4 selectively labels the tumor stroma, e.g., the cancer-associated fibroblast compartment. Conversely, MCT1 was selectively expressed in the epithelial cancer cells within the same tumors. Functionally, we show that overexpression of MCT4 in fibroblasts protects both MCF7 cancer cells and fibroblasts against cell death, under co-culture conditions. Thus, we provide the first evidence for the existence of a stromal-epithelial lactate shuttle in human tumors, analogous to the lactate shuttles that are essential for the normal physiological function of muscle tissue and brain. These data are consistent with the “reverse Warburg effect,” which states that cancer-associated fibroblasts undergo aerobic glycolysis, thereby producing lactate, which is utilized as a metabolic substrate by adjacent cancer cells. In this model, “energy transfer” or “metabolic-coupling” between the tumor stroma and epithelial cancer cells “fuels” tumor growth and metastasis, via oxidative mitochondrial metabolism in anabolic cancer cells. Most importantly, our current findings provide a new rationale and novel strategy for anti-cancer therapies, by employing MCT inhibitors.     

Introduction: why do we need a science of well-being?

Introduction: why do we need a science of well-being?     

Maintenance of polymorphic females: do parasites play a role?

The role of parasites in explaining maintenance of polymorphism is an unexplored research avenue. In odonates, female-limited color polymorphism (one female morph mimicking the conspecific male and one or more gynochromatic morphs) is widespread. Here we investigated whether parasitism contributes to color polymorphism maintenance by studying six species of female dimorphic damselflies using large databases of field-collected animals. We predicted that androchrome females (male mimics) would be more intensively parasitized than gynochrome females which is, according to previous studies, counterbalanced by the advantages of the former when evading male harassment compared to gynochrome females. Here we show that in Ischnura denticollis and Enallagma novahispaniae, androchrome females suffer from a higher degree of parasitism than gynochromatic females, and contrary to prediction, than males. Thus, our study has detected a correlation between color polymorphism and parasitic burden in odonates. This leads us to hypothesize that natural selection, via parasite pressure, can explain in part how androchrome and gynochrome female color morphs can be maintained. Both morphs may cope with parasites in a different way: given that androchrome females are more heavily parasitized, they may pay a higher fecundity costs, in comparison to gynochrome females.     

Deciding on a new home: how do honeybees agree?

A swarm of honeybees (Apis mellifera) is capable of selecting one nest-site when faced with a choice of several. We adapt classical mathematical models of disease, information and competing beliefs to such decision-making processes. We show that the collective decision may be arrived at without the necessity for any bee to make any comparison between sites.     

Age-related macular degeneration: what can a family physician do?

Age-related macular degeneration (ARMD) is the most common cause of blindness in Canada, and, as the name suggests, its incidence increases rapidly with age. Atrophic maculopathy, one of the forms of ARMD, is associated with only mild to moderate visual loss but is not treatable. On the other hand, exudative maculopathy, another form, is characterized by the formation of neovascular membranes and causes acute visual disturbances; however, it is potentially treatable by means of laser photocoagulation. In symptomatic patients blood, lipids or serous fluid seen on funduscopic examination indicates occult neovascularization, even if no neovascular membrane is visible. In asymptomatic patients their age, the presence of soft drusen and the coarse pigmentary disturbance have the greatest correlation with the risk of neovascularization. Suitability for laser photocoagulation depends on the stage of exudative maculopathy, the duration of symptoms and the patient''s visual acuity. Therefore, family physicians must know what the earliest symptoms are and how to identify patients at highest risk so that intervention in terms of education, screening and prompt referral to an ophthalmologist for confirmation and subsequent treatment can be instituted.     

Reperfusion-induced arrhythmias: do free radicals play a critical role?

This article assesses whether oxygen-derived free radicals are one of the molecular causes of life-threatening arrhythmias that arise upon reperfusion of the ischemic myocardium. Evidence supporting this proposition has been obtained from studies of the effects of free radical scavengers and antioxidants, free radical generating systems, inhibition of various sources of free radicals and studies investigating the formation of free radicals and their products during early reperfusion. It has been hypothesized that free radical formation causes localised membrane damage to the sarcolemma that results in focal alterations in transmembrane ionic fluxes, particularly potassium. These changes in ionic fluxes may then lead to electrophysiological abnormalities that culminate in ventricular arrhythmias.     

Plant biology: do DELLAs do defence?

DELLA proteins repress plant growth and developmental processes. Recent data suggest that DELLAs improve survival by imposing growth restraint during plant stress, enabling limited resources to be diverted to pathogen defence.