Increased physical activity may be more protective for metabolic syndrome than reduced caloric intake. An analysis of estimated energy balance in U.S. adults: 2007-2010 NHANES.

BACKGROUND AND AIM: The purpose of this study was to examine the association between physical activity (PA), caloric intake, and Metabolic Syndrome (MetS) in a representative sample of the United States population. METHODS AND RESULTS: Data for 4327 adults from 2007 to 2010 NHANES were analyzed. MetS was defined using both ATPIII and AHA/NHLBI criteria. Weekly moderate and vigorous physical activity (PA) minutes from work, leisure-time, and transportation PA were used to estimate Total Energy Expenditure (TEE) from Basal Metabolic Rate (BMR) using the Harris-Benedict equation. Average total calories (KCAL) from two 24-h dietary recalls were used to compare energy intake and expenditure between subjects with and without MetS. An alpha of 0.05 was used to determine statistical differences. The age adjusted prevalence of MetS was 21.9% (95% CI 20.1-23.6) and 36.8% (34.7-39.0) using ATPIII and AHA/NHLBI criteria, respectively. The estimated population mean for KCAL/TEE was 0.83 (95% CI 0.82-0.84), and the mean for KCAL/BMR was 1.25 (95% CI 1.23-1.27). Subjects without MetS (MetS-) reported 36 ± 13 (ATPIII) and 45 ± 18 (AHA/NHLBI) more daily moderate PA minutes than subjects with MetS (MetS+). At each level of PA, MetS- consumed more calories relative to BMR and TEE than MetS+. For both normal and overweight adults, KCAL/BMR was higher for MetS- than MetS+. For all BMI groups, there were no differences between MetS- and MetS+ with respect to KCAL/TEE. Though MetS+ adults in either MetS criteria were generally less physically active, MetS- adults maintained a higher caloric intake relative to estimated energy needs. CONCLUSIONS: These results suggest energy needs may be distorted in Metabolic Syndrome and increased physical activity may be more protective than reduced caloric intake.

Polymorphonuclear neutrophils express the common acute lymphoblastic leukemia antigen.

Monoclonal antibodies J5, VIL-A1, and BA-3, known to react with the common acute lymphoblastic leukemia antigen (CALLA) were found to specifically stain normal human polymorphonuclear neutrophils (PMN). The antigen detected on PMN had a molecular weight (95,000-110,000 mol wt) close to that of CALLA (95,000-100,000 mol wt) and thus these surface membrane antigens are likely related, if not identical. The fluorescent staining intensity of PMN is comparable to that of CALLA-positive leukemic cells and the presence of PMN in patient samples could potentially produce false-positive results in diagnosis.

Configuration and expression of the T cell receptor beta chain gene in human T-lymphotrophic virus I-infected cells.

We studied the configuration and expression of the gene encoding the beta chain of the T cell receptor (TCR beta) in cell lines and primary tumor cells infected by the human T cell leukemia/lymphoma (lymphotrophic) virus type I (HTLV-I). Most of the cell lines and all the primary tumor cells showed rearrangement of the TCR beta gene, and in each case the rearrangement was distinct. The majority of cases examined were clonal with respect to a particular TCR beta gene rearrangement. Primary tumor cells from one case (SD) were found to have a tandem duplication of a portion of chromosome 7; this appears to have resulted in the presence of three alleles of the TCR beta gene, each of which is arranged differently. This suggests that the chromosomal abnormality, and possibly infection by HTLV-I, occurred before TCR beta gene rearrangement. Cell lines infected by HTLV-I express levels of TCR beta mRNA similar to PHA stimulated lymphocytes, suggesting that this gene is not transcriptionally activated as a result of infection by HTLV-I. Cloned T cells of known antigen specificity that are infected by HTLV-I in vitro show impairment of immune function, including loss of antigen-specific responsiveness and the acquisition of alloreactivity. Comparison of the configuration of the TCR beta gene before and after infection revealed no changes detectable by Southern blot analysis. Levels of expression of the TCR beta gene at the mRNA level and surface expression of the T3 complex were also not significantly altered, suggesting that changes in immune function cannot be attributed to quantitative changes in the TCR molecule. The configuration of the TCR beta gene in primary tumor cells infected by HTLV-I was compared with that in the derived cell lines. In all pairs examined, the configuration in the primary tumor cells was different from that in the cell lines, strongly suggesting that the cells that grow in culture are not the original neoplastic cells.

Role of the thymus in directing the development of a subset of B lymphocytes.

In an effort to evaluate the role of the thymus in influencing the development of Lyb-5- B lymphocytes, mice expressing both the xid and nu gene defects were studied. Mice expressing either of these defects respond to both trinitrophenylated Brucellus abortus and lipopolysaccharide; whereas mice with the combined defect show markedly suppressed responses. The other abnormalities included: (a) greater than 80 percent diminution of serum Ig levels; (b) significant increase in the number of sIgM+ sIgD- B lymphocytes; (c) reduced expression of IgD on sIgD+ cells; and (d) a strikingly abnormal histology of their lymphoid tissue. Because nu/nu mice that do not express the xid defect appear relatively normal, it would suggest that the development of Lyb-5- B lymphocytes require a thymic influence for normal maturation, whereas, Lyb-5+ B lymphocytes are relatively independent of such influences.

Involvement of the bcl-2 gene in human follicular lymphoma.

Recombinant DNA probes were cloned for the areas flanking the breakpoint on chromosome 18 in cells from a patient with acute lymphocytic leukemia of the B-cell type; cells of this line carry the t(14;18) chromosomal translocation. Two of the probes detected DNA rearrangements in approximately 60 percent of the cases of follicular lymphoma screened. In follicular lymphoma, most of the breakpoints in band q21 of chromosome 18 were clustered within a short stretch of DNA, approximately 2.1 kilobases in length. Chromosome 18-specific DNA probes for the areas flanking the breakpoints also detected RNA transcripts 6 kilobases in length in various cell types. The gene coding for these transcript (the bcl-2 gene) seems to be interrupted in most cases of follicular lymphomas carrying the t(14;18) chromosomal translocation.

Functional properties of antigen-specific T cells infected by human T-cell leukemia-lymphoma virus (HTLV-I).

Tetanus-toxoid specific helper-inducer T-cell clones, which had been infected and transformed by human T-cell leukemia-lymphoma virus (HTLV-I), were obtained from an antigen-specific human T cell line by using a limiting dilution technique in the presence of the virus. These HTLV-I-infected T-cell clones proliferated specifically in response to soluble tetanus toxoid but, unlike normal T cells, they could do so in the absence of accessory cells. The HTLV-I-infected T-cell clones did not present the antigen to autologous antigen-specific T cells that were not infected with HTLV-I. The capacity of helper-inducer T cells to retain antigen-specific reactivity after infection by HTLV-I, while losing the normal T-cell requirement for accessory cells, has clinical and theoretical implications.

The t(14;18) chromosome translocations involved in B-cell neoplasms result from mistakes in VDJ joining.

In this study, the joining sequences between chromosomes 14 and 18 on the 14q+ chromosomes of a patient with pre-B-cell leukemia and four patients with follicular lymphoma carrying a t(14;18) chromosome translocation were analyzed. In each case, the involved segment of chromosome 18 has recombined with the immunoglobulin heavy-chain joining segment (JH) on chromosome 14. The sites of the recombination on chromosome 14 are located close to the 5' end of the involved JH segment, where the diversity (D) regions are rearranged with the JH segments in the production of active heavy-chain genes. As extraneous nucleotides (N regions) were observed at joining sites and specific signal-like sequences were detected on chromosome 18 in close proximity to the breakpoints, it is concluded that the t(14;18) chromosome translocation is the result of a mistake during the process of VDJ joining at the pre-B-cell stage of differentiation. The putative recombinase joins separated DNA segments on two different chromosomes instead of joining separated segments on the same chromosome, causing a t(14;18) chromosome translocation in the involved B cells.

Disruption of the human SCL locus by "illegitimate" V-(D)-J recombinase activity.

A fusion complementary DNA in the T cell line HSB-2 elucidates a provocative mechanism for the disruption of the putative hematopoietic transcription factor SCL. The fusion cDNA results from an interstitial deletion between a previously unknown locus, SIL (SCL interrupting locus), and the 5' untranslated region of SCL. Similar to 1;14 translocations, this deletion disrupts the SCL 5' regulatory region. This event is probably mediated by V-(D)-J recombinase activity, although neither locus is an immunoglobulin or a T cell receptor. Two other T cell lines, CEM and RPMI 8402, have essentially identical deletions. Thus, in lymphocytes, growth-affecting genes other than immune receptors risk rearrangements.

Drug development and the FDA's Critical Path Initiative.

Advances in biomedical research over recent decades have substantially raised expectations that the pharmaceutical industry will generate increasing numbers of safe and effective therapies. However, there are warning signs of serious limitations in the industry's ability to effectively translate biomedical research into marketed new therapies. Clinical pharmacologists should be aware of these signals and their potential impact. Here, we discuss a strategy, where clinical pharmacology can play an important role to improve the process of drug development.

Clonality of angioimmunoblastic lymphadenopathy and implications for its evolution to malignant lymphoma.

To investigate the relationship of the lymphoid hyperplasia of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) to supervening malignant lymphoma, we subjected DNA from lymph nodes and peripheral blood mononuclear cells from five AILD patients to Southern blot analysis to detect clonal rearrangements of immunoglobulin and T-cell receptor genes. Lymph nodes and peripheral blood from AILD patients were found to contain clones of lymphoid cells harboring either immunoglobulin or T-cell receptor gene rearrangements that, in some instances, regressed during the course of disease. A lymph node from one patient was involved by immunoblastic lymphoma and manifested an additional gene rearrangement pattern not seen in premalignant specimens from that patient. In contrast, DNA obtained from normal peripheral blood mononuclear cells and 11 examples of other forms of lymphoid hyperplasia showed no gene rearrangements. As a disorder of cellular immunoregulation in which lymphoid cells may escape normal growth controls, AILD provides a natural model to dissect stages of lymphomagenesis in man.

In vitro enhancement of immunoglobulin gene expression in chronic lymphocytic leukemia.

B cell chronic lymphocytic leukemia (CLL) cells appear to be arrested in their differentiation so that little immunoglobulin is secreted in most cases. To determine their capacity for further differentiation we stimulated cells from a series of 10 cases of CLL with a phorbol ester and assayed for production of immunoglobulin protein, accumulation of immunoglobulin mRNA, and alterations in cell surface markers. We found that cells from all cases were induced to secret monoclonal immunoglobulin of the same heavy and light chain type as the surface membrane immunoglobulin type. Immunoglobulin secretion was preceded by a rapid increase in the levels of mRNA coding for IgM, predominantly the secretory form, mu s-mRNA, rather than the membrane form, mu m-mRNA. A similar selection of mu s- over mu m-mRNA is known to occur in plasma cells by a mechanism of differential processing of mRNA from a single mu-chain gene. Except for a decline in the expression of surface IgD, cell surface determinants remained unaffected both in terms of the percentage of positive cells and the relative number of sites per cell. In contrast to previous studies, these results indicate that CLL cells consistently retain the capacity to further differentiate toward plasma cells and secrete immunoglobulin. The immunoglobulin secretion is mediated, at least in part, by a developmentally regulated increment in mu s-mRNA.

Infection of human T lymphotropic virus-I-specific immune T cell clones by human T lymphotropic virus-I.

Human T lymphotropic virus-I (HTLV-I)-specific T cell lines were established and cloned. K5, an OKT8+ clone bearing multiple proviral integration sites, retained its HTLV-I-specific cytotoxicity and a normal dependence on interleukin 2 (IL-2), indicating that there is a finite number of transforming integration sites. R2, an OKT4+ HTLV-I-infected clone, initially mounted a proliferative response to HTLV-I; but then its IL-2-independent proliferation increased and the antigen specificity was lost. All HTLV-I-infected clones tested including K7, another OKT8+ transformed cytotoxic clone that had lost its reactivity, expressed comparable levels of T cell receptor beta-chain (TCR-beta) messenger (m)RNA. Although clones K5 and K7 had different functional properties, they had the same rearrangement of the TCR-beta gene, suggesting that they had the same clonal origin. These data indicate that HTLV-I-specific T cells retain their immune reactivity for variable periods of time following infection, but then usually lose it; in some cases, however, no alteration in function can be detected. The data also suggest that different consequences can take place in the same clone depending on the pattern of retroviral infection.

Involvement of the bcl-2 gene in Hodgkin's disease.

A major obstacle to investigations of Hodgkin's disease is the paucity of malignant cells, i.e., Reed-Sternberg cells and their variants, in tissues of patients with this disease. Consequently, the pathogenesis, cell of origin, and clonality of this relatively frequent lymphoma have remained unresolved. Results of recent studies suggest that in some instances Reed-Sternberg cells carry rearranged immunoglobulin heavy-chain joining region (JH) loci as well as chromosomal translocations involving band 14q32. Prompted by these findings, we sought to determine if the t(14;18) (q32;q21) translocation of follicular, non-Hodgkin's B-cell lymphoma was associated with Hodgkin's disease. To detect the possible t(14;18) (q32;q21) translocation within the rare malignant cells of Hodgkin's disease, we amplified sequences created by the t(14;18) translocation using the polymerase chain reaction (PCR). With this approach, DNA sequences carrying the direct fusion of the major breakpoint region of the candidate oncogene, bcl-2, derived from chromosome 18q21, with JH on chromosome 14q32 can be detected in as few as one in 10(5)-10(6) cells. In the present study, joined bcl-2/JH sequences were detected in tissues involved by Hodgkin's disease in 17 of 53 (32%) patients. The frequent association of bcl-2 translocation with Hodgkin's disease suggests that this oncogene has a role in the pathogenesis of Hodgkin's disease. That bcl-2 is involved in a major class of lymphoma in addition to follicular lymphoma implies a role for additional factors responsible for generating the two distinctive clinical and pathologic disease states.

Molecular pathways of adhesion in spontaneous rosetting of T-lymphocytes to the Hodgkin's cell line L428.

Spontaneous rosetting of T-lymphocytes to Reed-Sternberg cells has been observed both in vitro and in vivo but its molecular mechanism has not been defined. We have investigated such rosetting using the Hodgkin's cell line L428. L428 expresses high levels of LFA-3 and ICAM-1, both of which are ligands for T-cell adhesion. Monoclonal antibody inhibition of spontaneous rosetting indicated that it is not dependent on the T-cell receptor complex but is largely mediated by interaction of T-cell CD2 (T11/E-rosette receptor) with its ligand LFA-3 on L428 cells. Studies using an alternate assay of adhesion (conjugate formation) confirm the roles of CD2/LFA-3 and also implicate a second mode of binding via LFA-1 on T-cells to ICAM-1 on L428. These data explain the previously reported finding of T-cell rosetting with Reed-Sternberg cells as an exaggeration of normal antigen-independent T-cell adhesion.

Clonal evolution of t(14;18) follicular lymphomas demonstrated by immunoglobulin genes and the 18q21 major breakpoint region.

A 2.8-kilobase major breakpoint region on chromosome segment 18q21 is the site of most t(14;18) translocations typical of human follicular lymphomas. Breaks are focused at the 5' end of joining (JH) regions of immunoglobulin (Ig) on chromosome 14, indicating that the translocation occurs at a pre-B-cell stage during attempted heavy (H) chain joining. A new gene from 18q21 (Bcl-2) is placed in the H chain locus creating a unique, translocation-specific JH;18q21 rearrangement that presumably represents a transformation event. In addition, normal Ig gene joining occurs in a H before light (L) chain and K before lambda cascade, creating ordered clonal markers. These serial markers were examined to determine if variations in Ig gene patterns during the natural history of lymphomas represent the emergence of truly separate neoplasms or heterogeneity of a single neoplasm. We examined 45 serial biopsies from 16 B follicular lymphoma patients; six cases showed variation in Ig gene patterns over time. Seven individuals had a detectable JH;18q21 rearrangement present, and it remained unchanged over 5-10 years. Further rearrangements of H chain genes occurred on the normal chromosome 14 within evolving subclones of the original tumor. Lambda L chains also underwent additional rearrangements in two instances, while K gene patterns remained unchanged. All variations in the normal H and L chain genes were 2 degrees rearrangements occurring at a mature B-cell stage following the initial successful rearrangement of a H and L chain. In contrast the t(14;18) breakpoint was conserved in each individual, indicating that evolving neoplastic subpopulations arose from a common clonal progenitor cell.

bcl-2 protein inhibits etoposide-induced apoptosis through its effects on events subsequent to topoisomerase II-induced DNA strand breaks and their repair.

Previous studies have shown that bcl-2 overexpression can inhibit apoptosis induced by DNA-damaging agents widely used in cancer chemotherapy, including X-irradiation, alkylating agents (hydroperoxycyclophosphamide, etc.), and topoisomerase II inhibitors (etoposide, etc.). However, little is known about the mechanism by which bcl-2 overexpression inhibits apoptosis triggered by these agents. In this study, we examined whether bcl-2 overexpression could have effects on etoposide-induced DNA damage and its repair. For these experiments, we developed CH31 clones (mouse B-cells) stably transfected with human bcl-2 sense plasmids and compared these clones with a parental CH31 clone or CH31 clones with antisense plasmids. Overexpression of bcl-2 protein inhibited etoposide-induced apoptosis and cytotoxicity. However, there was no or little difference in the production and repair of DNA-protein cross-links, DNA single-strand breaks, and double-strand beaks among a parental CH31 clone and CH31 clones with human bcl-2 sense or antisense plasmids. These findings indicate that (a) apoptosis or cytotoxicity induced by etoposide can be separated into early events (formation of double-strand breaks, DNA single-strand breaks, and double-strand breaks) and later events (secondary DNA fragmentation or cell death) and (b) bcl-2 inhibits apoptosis and cytotoxicity induced by etoposide at some steps between these events.

Transformation of Epstein-Barr virus immortalized human B-cells by chemical carcinogens.

Epstein-Barr virus-immortalized human lymphocytes were used to analyze the transition from the benign hyperproliferative to the malignant transformed state. Treatment with N-acetoxy-2-acetylaminofluorene, a potent frameshift mutagen, induced conversion of the Epstein-Barr virus immortalized lymphocytes into high-grade "immunoblastic lymphomas" on injection into athymic mice, whereas injection of the untreated, original cells did not. The tumor cells were all of the B-cell lineage as determined by the presence of surface immunoglobulins and antigens detected by B-cell specific antibodies to B1 and B4, and the absence of the T-cell-specific markers, 3A1 and LEU-1. The N-acetoxy-2-acetylaminofluorene-induced tumor lines displayed abnormal diploid to tetraploid karyotypes. The fewest chromosomal rearrangement, excluding tetraploidy, observed in these chemically induced lymphomas involved a deletion in chromosome 6, and additions on both chromosomes 16 and 4. Neither major rearrangements nor amplifications were found for K-ras, H-ras, N-ras, c-myc, Blym, and c-myb in these tumor lines.

Diversity of immunological phenotypes of lymphoblastic lymphoma.

Eleven cases of lymphoblastic malignancy, presenting as lymphoma, were investigated for immunological and differentiation markers prior to the onset of therapy. Biopsy specimens exhibited the typical morphological features of lymphoblastic lymphoma (convoluted T-cell lymphoma). Intranuclear terminal deoxynucleotidyl transferase was detected in the neoplastic cells from each case by indirect antibody staining of cytocentrifuge preparations. Eight cases were T-cell type as evidenced by unsensitized sheep erythrocyte rosette formation and staining with the monoclonal antibody OKT11. Three T-cell cases were OKT4 positive, two were OKT8 positive, and none were positive with both OKT4 and OKT8. Three cases failed to react with any monoclonal antibodies specific for T-cells and did not form unsensitized sheep erythrocyte rosettes or stain for surface immunoglobulin. However, these three cases were Ia positive and J5 (common acute lymphoblastic leukemia antigen) positive. Cells from two of these erythrocyte rosette-negative, Ia-positive, common acute lymphoblastic leukemia-positive cases contained intracytoplasmic mu heavy chains and were therefore of pre-B-cell phenotype. These cases were histologically indistinguishable from the T-cell cases. However, clinically, they were distinguished by the absence of mediastinal masses and by a clinical presentation as isolated lytic lesions of bone in two of the three. OKT9 and OKT10 stained neoplastic cells from T-cell, as well as pre-B-lymphoblastic, lymphoma. Although morphologically homogeneous, lymphoblastic lymphomas are comprised of an immunologically diverse group of neoplasms which include cells of "common" and "mature" thymocyte, non-T, non-B, and pre-B phenotypes and are closely related to the cells of acute lymphoblastic leukemia. In addition, intratumor heterogeneity was observed in most instances and may reflect growth or differentiation differences between subpopulations of individual neoplastic clones.

Objective regressions of T- and B-cell lymphomas in patients following treatment with anti-thymocyte globulin.

We have conducted a clinical trial utilizing anti-thymocyte globulin (ATG) for the treatment of patients with non-Hodgkin's lymphomas. Six patients were treated; 50% reductions in tumor mass of short duration were observed in one patient with a T-cell lymphoma and two patients with B-cell lymphomas. In vitro assays have been performed in an attempt to study the reactivity and potential mechanism of antitumor action of the ATG. The ATG bound to essentially all normal blood mononuclear leukocytes as well as tumor cells from patients with T-, B-, or null cell lymphomas demonstrating its lack of specificity. Furthermore, complement-mediated lysis of normal mononuclear leukocytes, normal T- or B-cells, and tumor cells from two unresponsive patients were all comparable; moreover, since this lysis occurred only at concentrations of ATG that are not attainable in vivo, it is unlikely that complement-mediated cytotoxicity accounts for the responses observed. Peripheral blood lymphocyte counts and total erythrocyte rosettes did decrease during ATG treatment. Thus, objective tumor responses in both B- and T-cell non-Hodgkin's lymphomas can be achieved with a very nonspecific antiserum although significant toxicity resulted. Whether the magnitude or duration of response can be increased with monoclonal antibodies remains to be determined. Future success with serotherapy might require use of either a battery of different monoclonal antibodies or a single monoclonal antibody that can deliver radioisotopes, chemotherapy, or toxins to the tumor cells.