Lourdes Domínguez-Gerpe

Identification of molecular mechanisms related to nonthyroidal illness syndrome in skeletal muscle and adipose tissue from patients with septic shock

Objective  Septic shock is one of various causes of nonthyroidal illness syndrome (NTIS). In humans, the molecular mechanisms involved in NTIS are mostly unknown. The aim of this study was to investigate, in patients with NTIS secondary to septic shock, changes in the expression of genes involved in the actions of thyroid hormones and in the activity of deiodinase enzymes, in two tissues important for protein and energy metabolism, skeletal muscle (SM) and subcutaneous adipose tissue (SAT).

Alterations induced by chronic stress in lymphocyte subsets of blood and primary and secondary immune organs of mice

BackgroundThe immune system is particularly sensitive to stress. Although acute stress generally has positive effects, chronic stress typically provokes immunosuppression. The elucidation of the mechanisms involved in immunosuppression are of interest for the design of therapeutic approaches to avoid the appearance of stress disorders. This study aimed to investigate chronic stress-induced alterations on lymphocyte subset distribution and percentages. The experiments were performed with C57BL/6 mice subjected to chronic immobilization stress.ResultsStress caused a marked increase in apoptosis inside the thymus, and a reduction in the total number of thymocytes. Furthermore, the proportion of immature thymocytes declined significantly suggesting that the increased apoptosis mainly affected cells of immature phenotype. In blood, the total number of lymphocytes diminished but not all lymphocyte populations showed the same tendency: while the relative proportion of B cells declined slightly, the relative proportion of circulating CD3+ cells, and particularly some T cell subsets showing an immature phenotype (CD3+PNA+), increased under stress. The spleen and lymph nodes show a marked reduction in cellularity, but the relative proportion of T cells increased, while no change or only a slight reduction was observed in the relative proportion of B cells. Similarly, the relative proportion of T cells increased in bone marrow.ConclusionsDetailed data on the alterations of lymphoid cell subsets occurring under immobilization stress, both in the bloodstream and in different lymphoid tissues, are obtained. In general, T cells are more affected than B cells and, in particular, a marked increase in the percentage of a subset of circulating PNA+CD3+ T cells is observed.

Time-course of the murine lymphoid tissue involution during and following stressor exposure

Groups of 35-day-old male C57BL/6 mice were stressed 1 hour per day by immobilization for 1, 2, 3, 5, 8, 11 or 14 consecutive days. Control groups were left undisturbed. The animals were then killed and body weight and the weights of the thymus, spleen and axillary lymph nodes determined. Chronic immobilization stress caused involution of the thymus, spleen and lymph nodes to an extent depending on the number of days of stress. The thymus showed the fastest response: thymus weight was significantly lower in stressed animals than in controls by the third day of stress while significant effects on spleen and lymph node weight were not observed until day 5. Fast recovery of lymphoid organ weight was observed after the stress period. The thymus recovered most quickly: control values were re-attained approximately 8 days after cessation of stress, and indeed by day 20 thymus weight was about 12% higher than in normal animals. The spleen and lymph nodes recuperated weight more slowly, re-attaining control values after about 20 days.

Site-dependent differences in both prelamin A and adipogenic genes in subcutaneous adipose tissue of patients with type 2 familial partial lipodystrophy

Background: Type 2 familial partial lipodystrophy (FPLD2) is characterised by loss of fat in the limbs and buttocks and results from mutations in the LMNA gene. Aim: To evaluate the role of several genes involved in adipogenesis in order to better understand the underlying mechanisms of regional loss of subcutaneous adipose tissue (scAT) in patients with FPLD2. Methods: In total, 7 patients with FPLD2 and 10 healthy control participants were studied. A minimal model was used to calculate the insulin sensitivity (IS). scAT was obtained from abdomen and thigh by biopsy. Relative gene expression was quantified by real-time reverse transcription PCR in a thermal cycler. Prelamin A western blot analysis was carried out on scAT and prelamin A nuclear localisation was determined using immunofluorescence. Adipocyte nuclei were examined by electron microscopy. Results: Patients with FPLD2 were found to have significantly lower IS. The expression of LMNA was similar in both groups. The expression of PPARG2, RB1, CCND3 and LPL in thigh but not in abdomen scAT was significantly reduced (67%, 25%, 38% and 66% respectively) in patients with FPLD2. Significantly higher levels of prelamin A were found in peripheral scAT of patients with FPLD2. Defects in the peripheral heterochromatin and a nuclear fibrous dense lamina were present in the adipocytes of patients with FPLD2. Conclusions: In FPLD2 participants, prelamin A accumulation in peripheral scAT is associated with a reduced expression of several genes involved in adipogenesis, which could perturb the balance between proliferation and differentiation in adipocytes, leading to less efficient tissue regeneration.

A novel phenotypic expression associated with a new mutation in LMNA gene, characterized by partial lipodystrophy, insulin resistance, aortic stenosis and hypertrophic cardiomyopathy

Background  Lipodystrophies are a heterogeneous group of diseases characterized by abnormal fat distribution. Familial partial lipodystrophy 2 (FPLD2) is due to mutations in the LMNA gene. Previous studies have suggested that LMNA mutations 5′ to the nuclear localization signal (NLS) are more likely to underlie laminopathies with cardiac or skeletal muscle involvement, while mutations 3′ to the NLS are more likely to underlie lipodystrophy and progeroid syndromes.

Prematurely aged children: molecular alterations leading to Hutchinson-Gilford progeria and Werner syndromes.

Ageing is thought to be a polygenic and stochastic process in which multiple mechanisms operate at the same time. At the level of the individual organism ageing is associated with a progressive deterioration of health and quality of life, sharing common features such as: alopecia and grey hair, loss of audition, macular degeneration, neurodegeneration, cardiovascular diseases, osteoporosis, cataract formation, type-2 diabetes, lipodystrophies; a generally increased susceptibility to infection, autoimmune disorders and diseases such as cancer; and an impaired ability to cope with stress. Recent studies of mechanisms involved in the ageing process are contributing to the identification of genes involved in longevity. Monogenic heritable disorders causing premature ageing, and animal models have contributed to the understanding of some of the characteristic organism-level features associated with human ageing. Werner syndrome and Hutchinson-Gilford progeria syndrome are the best characterized human disorders. Werner syndrome patients have a median life expectancy of 47 years with clinical conditions from the second decade of life. Hutchinson-Gilford progeria syndrome patients die at a median age of 11-13 years with clinical conditions appearing soon after birth. In both syndromes, alterations in specific genes have been identified, with mutations in the WRN and LMNA genes respectively being the most closely associated with each syndrome. Results from molecular studies strongly suggest an increase in DNA damage and cell senescence as the underlying mechanism of pathological premature ageing in these two human syndromes. The same general mechanism has also been observed in human cells undergoing the normal ageing process. In the present article the molecular mechanisms currently proposed for explaining these two syndromes, which may also partly explain the normal ageing process, are reviewed.

Age-Related Changes in Primary and Secondary Immune Organs of the Mouse

The present work describes the murine immune tissue evolution with age with special emphasis on the bone marrow. To that effect we monitored the weights of the thymus, spleen and axillary lymph nodes over the first year of life in C57BL/6 male and female mice. In addition, we monitored the relative proportions of erythroid, lymphoid and myeloid cells in the bone marrow, and performed in vitro migration assays of bone marrow cells to thymic supernatants, with the aim of determining whether the migration of such cells or the thymic attractive capacity are affected by age. Before puberty, a remarkable decline in the relative weight of the thymus, spleen and lymph nodes was observed; after that stage, however, only the thymus showed an involution. The proportion of myeloid cells in the bone marrow showed an increase with age. Furthermore, the migration of myeloid cells to thymic supernatants increased with age and paralleled the time-course of the myeloid cell increase found in the bone marrow. More interestingly, the proportion of lymphoid cells to total bone marrow cells showed a clear decline with age. The time-course of this decline closely paralleled that of thymus weight, suggesting that the involution of the thymus may be related to changes in the cell composition of the bone marrow.

Modulation of stress-induced murine lymphoid tissue involution by age, sex and strain : role of bone marrow

C57BL/6 and Balb C male and female mice of various ages were stressed by immobilization for 1 h/day (1, 2, 3, 5, 8, 11 or 14 consecutive days). The animals were then killed for determination of total body weight and the weights of the thymus, spleen and axillary lymph nodes. In addition, the total number of cells in the thymus and the proportion of lymphoid cells in the bone marrow cell population was defined. The effects of stress were modulated by age, sex and strain. Stress-induced involution of the thymus was generally more pronounced in older animals, while for the spleen was the opposite. Involution of the thymus was higher in males than in females, but there were no marked differences between the sexes in the response of the spleen. In general C57BL/6 mice were more sensitive to stress than Balb C mice. However, for the involution induced by stress on lymph nodes there were not a clear trend with age, sex or strain. In male and female mice of all ages and both strains, stress led to statistically significant reductions in the absolute number of cells inside the thymus and spleen and in the proportion of lymphoid cells in the bone marrow.

Prevalence and functional analysis of the S107P polymorphism (rs6647476) of the monocarboxylate transporter 8 (SLC16A2) gene in the male population of north-west Spain (Galicia).

Objective  Mutations in SLC16A2, the gene encoding the thyroid hormone (TH)‐specific transporter monocarboxylate transporter 8 (MCT8), result in a thyroid phenotype and severe mental retardation caused by neuronal TH deficiency. These mutational effects raise the question of whether polymorphic variation in SLC16A2 may also be associated with differences in serum levels of TH and/or TSH.

Lymphocyte protein synthesis: evidence that murine T cells are more affected by stress than B cells☆

B and T cells present in the spleen and other sites of the immune system play a crucial role in the protection of individuals by mounting a specific primary and secondary immune response. Disturbances in their signaling and functioning could lead to a deterioration of the defense mechanisms and thereby lead to infections, pathologies or diseases. In this work, we studied the effects of stress on the protein synthesis and metabolism of mouse splenocytes. The study was done by radiolabeling the entire mouse with 35S-methionine (in vivo procedure) or by culturing spleen cells under various conditions of stimulation in the presence of 35S-methionine (in vitro labeling). The stimulus was lipopolysaccharide (LPS), LPS + interleukin-4 (IL-4) and concanavalin A (on A). Samples from immobilization stressed and control animals were studied in parallel. The results showed minimum alterations due to stress on B cells, though a small decrease in proliferative capacity was observed. In contrast, T cells are profoundly affected by stress. More than 100 new proteins are expressed on 2D-gel patterns of T cells from stressed animals as compared with controls, some of which could be implicated in different signaling, or could have appeared because of an alteration in a pathway of synthesis, or even as a consequence of a change in the composition of T cell populations induced by stress.