Expression of glycolytic isoenzymes in activated human peripheral lymphocytes: cell cycle analysis using flow cytometry

Human peripheral lymphocytes activated with concanavalin A and phorbol myristate ester exhibit an increase in glycolysis on a time-course similar to that for DNA synthesis. Elevated glycolysis is accompanied by increased specific activities of the glycolytic enzymes. Increased enzyme activities are accounted for by the appearance of specific isoenzyme forms (muscle forms) normally expressed in rapidly growing tumor cells or in growth-stimulated cells. In the present study we analyzed the expression of the glycolytic isoenzymes during cell cycle progression of activated human lymphocytes using two-parameter (DNA and protein) flow cytometry. Time-course studies and analysis of subpopulations prepared by elutriation centrifugation showed that the inducible isoenzymes are expressed at low levels or not at all in G0 cells. They are expressed first during the G0 to G1 transition or in early G1. However, expression increases throughout G1, reaching a maximum in S-phase. Thus, induction of glycolytic isoenzymes provides an excellent marker of T-cell activation and progression toward DNA synthesis.     

Flow sorting of tumor cells for morphometric analysis, particularly of rare cells.

Using flow cytometric DNA measurement and sorting combined with morphometric light microscopy, different groups of cells were studied in a human melanoma pleural effusion, a human melanoma lymph node metastasis and a mouse tumor, as well as in normal reference tissues. Beside cells of the predominant tumor cell population, three types of rare tumor cells were studied after enrichment by sorting: a) giant cells from the greater than 8c region, comprising about 5% of the tumor cells, b) binucleated and multinucleated cells with unequal nuclear sizes within the same cell, found at frequencies of about 1.5%, and c) less than 2c cells which were derived from the so-called "debris"-region of the DNA histogram, found at frequencies of about 1 to 6%. All these rare cells were found only in the malignant tumors and not in the benign reference tissues. Morphometry showed that the increase in the cellular DNA content in the different fractions of tumor cells was combined with an increase in the cellular and nuclear sizes. However, the n/c-ratio was constant in the whole range of tumor cell fractions, including the fractions from the the less than 2c and the greater than 8c regions. The n/c-ratio of the less than 2c cells and giant cells differed from that of corresponding normal cells underlining their origin from the predominant tumor cell population. The possible linkage between the occurrence of the three rare cell types and genetic instability of tumors related to faulty nucleus and cell division is discussed.     

The dynamics of production and destruction: Analytic insight into complex behavior

This paper analytically explores the properties of simple differential-difference equations that represent dynamic processes with feedback dependent on prior states of the system. Systems with pure negative and positive feedback are examined, as well as those with mixed (positive/negative) feedback characteristics. Very complex time dependent behaviors may arise from these processes. Indeed, the same mechanism may, depending on system parameters and initial conditions, produce simple, regular, repetitive patterns and completely irregular random-like fluctuations.For the differential-delay equations considered here we prove the existence of: (i) stable and unstable limit cycles, where the stable cycles may have an arbitrary number of extrema per period; and (ii) chaos, meaning the presence of infinitely many periodic solutions of different period and of infinitely many irregular and mixing solutions.     

PKM2, cancer metabolism,and the road ahead

A major metabolic aberration associated with cancer is a change in glucose metabolism. Isoform selection of the glycolytic enzyme pyruvate kinase has been implicated in the metabolic phenotype of cancer cells, and specific pyruvate kinase isoforms have been suggested to support divergent energetic and biosynthetic requirements of cells in tumors and normal tissues. PKM2 isoform expression has been closely linked to embryogenesis, tissue repair, and cancer. In contrast, forced expression of the PKM1 isoform has been associated with reduced tumor cell proliferation. Here, we discuss the role that PKM2 plays in cells and provide a historical perspective for how the study of PKM2 has contributed to understanding cancer metabolism. We also review recent studies that raise important questions with regard to the role of PKM2 in both normal and cancer cell metabolism.     

Effect of land use on pollution status and risk of fish endocrine disruption in small farmland ponds

To study whether the intensity of agricultural activities affects pesticides loads in pond environment, a large number of Belgian farmland ponds were surveyed in spring 2004. Temporal distribution of pollutants was also investigated over restricted survey ponds sampled three times round year 2007. Sedentary pond Prussian carp juveniles were also captured to determine their brain aromatase activity (AA) and plasma vitellogenin (VTG) levels. Heavy metal distribution was also examined in various pond matrices. Amongst the pesticides analysed, only herbicides were detected. Contamination of pond water by atrazine was frequently observed during spring 2004, while isoproturon and glyphosate were detected round year 2007. Levels of herbicides were inversely related to the distance of ponds to crop field, and values peaked in April or October. Absence of endocrine disruptors in pond water was confirmed by lack of modulation in VTG and AA in male fish. Heavy metals were present in all the pond matrices, but overall contamination levels were low. The results demonstrated that Belgian ponds were mainly contaminated by herbicides and that pond sedentary fish were not affected by endocrine disruptors. They also demonstrated a marked effect of land-use intensity on herbicide pollution which can be mitigated by an adjustment of the buffer zones.     

ATP consumption promotes cancer metabolism

Cancer cells metabolize glucose by aerobic glycolysis, a phenomenon known as the Warburg effect. Fang et al. (2010) show that the endoplasmic reticulum enzyme ENTPD5 promotes ATP consumption and favors aerobic glycolysis. The findings suggest that nutrient uptake in cancer cells is limited by ATP and satisfies energy requirements other than ATP production.