A Brain Anti-Senescence Transcriptional Program Triggered by Hypothalamic-Derived Exosomal microRNAs.

In contrast to the hypothesis that aging results from cell-autonomous deterioration processes, the programmed longevity theory proposes that aging arises from a partial inactivation of a "longevity program" aimed at maintaining youthfulness in organisms. Supporting this hypothesis, age-related changes in organisms can be reversed by factors circulating in young blood. Concordantly, the endocrine secretion of exosomal microRNAs (miRNAs) by hypothalamic neural stem cells (htNSCs) regulates the aging rate by enhancing physiological fitness in young animals. However, the specific molecular mechanisms through which hypothalamic-derived miRNAs exert their anti-aging effects remain unexplored. Using experimentally validated miRNA-target gene interactions and single-cell transcriptomic data of brain cells during aging and heterochronic parabiosis, we identify the main pathways controlled by these miRNAs and the cell-type-specific gene networks that are altered due to age-related loss of htNSCs and the subsequent decline in specific miRNA levels in the cerebrospinal fluid (CSF). Our bioinformatics analysis suggests that these miRNAs modulate pathways associated with senescence and cellular stress response, targeting crucial genes such as Cdkn2a, Rps27, and Txnip. The oligodendrocyte lineage appears to be the most responsive to age-dependent loss of exosomal miRNA, leading to significant derepression of several miRNA target genes. Furthermore, heterochronic parabiosis can reverse age-related upregulation of specific miRNA-targeted genes, predominantly in brain endothelial cells, including senescence promoting genes such as Cdkn1a and Btg2. Our findings support the presence of an anti-senescence mechanism triggered by the endocrine secretion of htNSC-derived exosomal miRNAs, which is associated with a youthful transcriptional signature.

Long Range Endocrine Delivery of Circulating miR-210 to Endothelium Promotes Pulmonary Hypertension.

RATIONALE: Unproven theories abound regarding the long-range uptake and endocrine activity of extracellular blood-borne microRNAs into tissue. In pulmonary hypertension (PH), microRNA-210 (miR-210) in pulmonary endothelial cells promotes disease, but its activity as an extracellular molecule is incompletely defined. OBJECTIVE: We investigated whether chronic and endogenous endocrine delivery of extracellular miR-210 to pulmonary vascular endothelial cells promotes PH. METHODS AND RESULTS: Using miR-210 replete (wild-type [WT]) and knockout mice, we tracked blood-borne miR-210 using bone marrow transplantation and parabiosis (conjoining of circulatory systems). With bone marrow transplantation, circulating miR-210 was derived predominantly from bone marrow. Via parabiosis during chronic hypoxia to induce miR-210 production and PH, miR-210 was undetectable in knockout-knockout mice pairs. However, in plasma and lung endothelium, but not smooth muscle or adventitia, miR-210 was observed in knockout mice of WT-knockout pairs. This was accompanied by downregulation of miR-210 targets ISCU (iron-sulfur assembly proteins)1/2 and COX10 (cytochrome c oxidase assembly protein-10), indicating endothelial import of functional miR-210. Via hemodynamic and histological indices, knockout-knockout pairs were protected from PH, whereas knockout mice in WT-knockout pairs developed PH. In particular, pulmonary vascular engraftment of miR-210-positive interstitial lung macrophages was observed in knockout mice of WT-knockout pairs. To address whether engrafted miR-210-positive myeloid or lymphoid cells contribute to paracrine miR-210 delivery, we studied miR-210 knockout mice parabiosed with miR-210 WT; Cx3cr1 knockout mice (deficient in myeloid recruitment) or miR-210 WT; Rag1 knockout mice (deficient in lymphocytes). In both pairs, miR-210 knockout mice still displayed miR-210 delivery and PH, thus demonstrating a pathogenic endocrine delivery of extracellular miR-210. CONCLUSIONS: Endogenous blood-borne transport of miR-210 into pulmonary vascular endothelial cells promotes PH, offering fundamental insight into the systemic physiology of microRNA activity. These results also describe a platform for RNA-mediated crosstalk in PH, providing an impetus for developing blood-based miR-210 technologies for diagnosis and therapy in this disease.

Analysis of the contribution of nonresident progenitor cells and hematopoietic cells to reparative dentinogenesis using parabiosis model in mice.

INTRODUCTION: The aim of this study was to analyze the contribution of nonresident progenitor/stem cells and hematopoietic cells to reparative dentinogenesis. METHODS: Parabiosis was established between C57BL/6-TgN(ACTbEGFP)10sb/J transgenic mice (GFP+) and C57BL/6 wild-type mice (GFP-) to ensure blood cross-circulation between animals. Reparative dentinogenesis was stimulated by pulp exposures and capping on the first maxillary molar in the GFP- mice. Histologic sections of injured molars from GFP- mice were analyzed by epifluorescence microscopy to examine the contributions of GFP+ cells (nonresident progenitor cells and hematopoietic cells originating from GFP+ mice) to reparative dentinogenesis. RESULTS: GFP+ cells were detected in close association with reparative dentin formed at the site of pulp exposure in the maxillary first molars of the GFP- mice. CONCLUSIONS: The present study suggests the participation of the nonresident progenitor cells and hematopoietic cells in reparative dentinogenesis.

Teorias sobre a atuacao dos medicamentos homeopaticos / Theories about the actuation of the homeopathic drugs

Correlacionando teorias, discutiremos aspectos da Homeopatia no sentido de mostrar a importancia de dar-lhe uma base teorica cientificae, como os ensinamentos da neurofisiologia cortico-visceral de Pavlov podem lhe ser uteis, principalmente na explicacao do modo de acao dos medicamentos homeopaticos no organismo

Teorias sobre a atuacao dos medicamentos homeopaticos / Theories about the actuation of the homeopathic drugs

Correlacionando teorias, discutiremos aspectos da Homeopatia no sentido de mostrar a importancia de dar-lhe uma base teorica cientificae, como os ensinamentos da neurofisiologia cortico-visceral de Pavlov podem lhe ser uteis, principalmente na explicacao do modo de acao dos medicamentos homeopaticos no organismo

Experimental studies on a lethal gene (t) in the Mexican axolotl, Ambystoma mexicanum.

In the Mexican axolotl, Ambystoma mexicanum, the developmental mutation lethal t is inherited as a simple Mendelian recessive. Mutant larvae failed to feed and died, on the average, 17 days after hatching. Unfed wild-type larvae died an average of 23 days after hatching. By 15 days, forelimb development had progressed further in the wild type; a cartilaginous scapula and humerus were present, but no cartilage was seen in the mutant limb. Histological examination indicated that the visceral cartilage may also be abnormal, and the rectus cervicus muscle was found to have fewer and smaller fibers. Though the mutant was not rescued by parabiosis with wild-type embryos, transplants of presumptive gill and limb tissue to wild-type hosts survived, indicating that the mutation is not an autonomous cell lethal.

Evidence that the premature death mutation (p) in the Mexican axolotl (Ambystoma mexicanum) is not an autonomous cell lethal.

Cell-lethal developmental mutations, which are presumed to affect the viability of all cells in a mutant embryo, have been distinguished from other development lethals on the basis of the results of parabiosis and transplant experiments. Premature death (p), previously classified as a cell lethal, does not survive parabiosis. However, transplants involving mutant eye, flank epidermis and primordial limb tissue all survived on a normal recipient. The mutant, therefore, cannot be considered a true cell lethal, though it suffers from serious and widespread abnormalities that cannot be corrected by parabiosis. In addition, transplants of mutant branchial mound tissue did not develop into normal gills on a normal recipient. These transplants were the only ones involving mutant endoderm, and their failure supports our hypothesis that the mutation leads to a specific endoderm defect.

Genetic and experimental studies on three associated mutant genes in the Mexican axolotl: st (for stasis), mi (for microphthalmic) and h (for hand lethal).

Three mutant genes, st, mi, and h, were discovered in an axolotl male received from Mexico City. All three are recessive to their normal alleles, and appear to segregate independently. Larvae homozygous for st (for stasis) suffer blockage of the circulation at hatching or shortly after, and the majority soon die; any surviving live only a few weeks at most. The mi/mi (microphthalmic) can be identified at the feeding stage. None survives more than a few days. The h/h (hand lethals) live until the digits have appeared on the forelimb. They may then be recognized by the thumb-like orientation of digit 1. Transplants from st/st embryos into normal produce normal structures which persist indefinitely. Those from mi/mi and h/h donors do not survive. The structures (forelimb, gills) derived from h/h donors grow for a relatively long time, and their final death and degeneration result in defects leading to death of most of the recipients. Parabiosis is of no benefit to st/st or h/h mutants and leads to the death of the normal twin; mi/mi mutants undergo a gradual absorption by the normal twin.

Experimental studies on a mutant gene (p) causing premature death of Ambystoma mexicanum embryos.

The premature death (p) mutation is a recessive lethal, which, in the homozygous condition, gives rise to a complex of abnormalities. The mutant embryos develop only to stage 37, at which time disintegration of superficial tissue begins. Many of the abnormalities observed in sections of the stage-37 mutant embryo are related to its failure to establish a functioning circulatory system, or to the resulting edema and/or ascites that distend the abdomen and flanks. There are, however, abnormalities of heart, liver, gill and muscle development which cannot be attributed to lack of circulation and edema. All of these abnormalities can be indirectly related to the endoderm, particularly the anterior and dorsal endoderm. The findings, therefore, suggest that the mutation leads to a fairly general defect of the endoderm.

Experimental studies on a lethal gene (1) in the Mexican axolotl, Ambystoma mexicanum.

1. Gene l is a recessive lethal factor found in the white strain of axolotls. Animals heterozygous for the gene are phenotypically normal. When mated with each other they give offspring 25% of which exhibit the lethal effects of the gene. 2. The l/l homozygotes develop normally to an advanced embryonic stage (Harrison stage 40) before the effects of the gene are first manifested. They then come to display a characteristic combination of abnormalities, including a disproportionately small head, small and poorly developed eyes, abnormal poorly developed gills, undifferentiated limb buds, and reduced overall growth rate. They may feed briefly, but soon stop and invariably die within a few weeks of the time of hatching. 3. The action of gene l has been analyzed by parabiosing mutant and normal embryos, and by grafting various organ primordia reciprocally between mutant and normal embryos. Parabiosis to normal embryos fails to correct the abnormalities of the mutants, although their survival may be somewhat prolonged. Grafts of mutant organ primordia (eye, limb, gill, pronephros, gonad, head) also invariably fail to show improved development or to survive on normal hosts; normal organ primordia develop normally on mutant hosts so long as the mutant survives. These experiments indicate that gene l is a recessive autonomous cell lethal affecting all of the organ systems during late embryonic and early larval development.